Oxygen reduction disposable kits, devices and methods of use thereof

ABSTRACT

The present disclosure relates to Oxygen Reduction Disposable kits (ORDKit), devices and methods for the improved preservation of whole blood and blood components. The improved devices and methods for the collection of blood and blood components provide for whole blood and blood components having reduced levels of oxygen. The devices and methods provide for the rapid preparation of deoxygenated blood and blood components for storage that improves the overall quality of the transfused blood and improves health outcomes in patients.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/052,738, filed Aug. 2, 2018 (allowed), which is a continuation ofU.S. application Ser. No. 15/459,813, filed Mar. 15, 2017, now U.S. Pat.No. 10,058,091, which is a continuation of PCT Application No.PCT/US2016/021794, filed Mar. 10, 2016, wherein InternationalApplication No. PCT/US2016/021794 claims the benefit of U.S. ProvisionalApplication No. 62/131,130, filed Mar. 10, 2015 and the contents of eachof which are herein incorporated by reference in their entireties.

FIELD OF THE INVENTION

The present disclosure relates to Oxygen Reduction Disposable kits(ORDKit), devices and methods for the improved preservation of wholeblood and blood components. More particularly, the disclosure relates tothe improved devices and methods for the collection of blood and bloodcomponents to provide whole blood and blood components having reducedlevels of oxygen. The methods, devices and kits of the presentdisclosure provide for improved quality of blood and blood componentsfor transfusion and improved patient safety and outcome.

BACKGROUND OF THE INVENTION

The supplies of liquid blood and blood components are currently limitedby storage systems used in conventional blood storage practices. Usingcurrent systems, stored blood expires after a period of about 42 days ofrefrigerated storage at a temperature above freezing (i.e., 4° C.) aspacked blood cell preparations. For Example, the World HealthOrganization (WHO) estimates more than 100 million units of blood arecollected and stored globally each year. In the US alone, there were13.6 million units of red blood cells (RBCs) collected in 2013 accordingto the American Association of Blood Bankers. During refrigeratedstorage, RBCs become progressively damaged by storage lesions. Whentransfused within the current 6-week limit, stored RBCs have lowerquality as well as potential toxicity, which can be manifested as sideeffects of transfusion therapy. Among the observed storage lesions arealtered biochemical and physical parameters associated with stored redblood cells. Examples of these alterations include in vitro measuredparameters such as reduced metabolite levels (adenosine triphosphate(ATP) and 2,3 diphosphoglycerate (2,3-DPG)), increased levels ofcell-free iron, hemolysis, increased levels of microparticles, reducedsurface area, echinocytosis, phosphatidylserine exposure, and reduceddeformability. Expired blood cannot be used and must be discardedbecause it may harm the ultimate recipient. These reasons and otherslimit the amount of readily available high quality blood needed fortransfusions.

When stored conventionally, stored blood undergoes a steadydeterioration which is associated with hemolysis, hemoglobin degradationand reduced ATP and 2,3-DPG concentrations. When transfused into apatient, the effects of the steady deterioration during storagemanifest, for example, as a reduction in the 24-hour in vivo recovery.Red blood cells stored for an extended period of time under conventionalconditions deteriorate and up to 25% may be removed by the recipient'sbody shortly after transfusion. Non-viable RBCs cause iron overload inchronically transfused patients. Hemoglobin in RBCs does not releaseoxygen efficiently at tissues due to loss of 2,3-DPG. RBCs are not ableto enter and perfuse capillary beds due to loss of deformability.Storage lesions in transfused blood may lead to major organ failure inthe lungs, heart, kidney, liver, and central nervous system, amongothers. Storage lesions in transfused blood may be associated withincreased morbidity.

Transfusing RBCs stored under conventional conditions for longer periodsmay result in higher morbidity and longer hospital stays compared totransfusing “fresher” red cells. Higher morbidity and longer hospitalstays result with RBCs that are stored longer than 3 weeks, incomparison to fresher red cells. For example, negative clinical outcomesin cardiac surgery occur when using “older” blood, multiple organfailure in surgical patients is related to the age of transfused redcells, correlations exist between older units and increased mortality insevere sepsis, failure to improve O₂ utilization is attributed todecreased 2,3-DPG, and decreased cardiac index is associated withincreased blood viscosity.

In addition to immediate removal by the recipient of certain RBCs,consequences of RBC storage lesions include: (i) depletion of ATP (lossof RBC's ability to dilate the pre-capillary arteriole); (ii) depletionof 2,3-DPG; (iii) accumulation of oxidative damage caused by reactiveoxygen species (ROS) formed by the reaction of denatured hemoglobin withO₂; and (iv) decreased RBC deformability and increased RBC viscosity,caused in part by oxidative damage to membrane and cytoskeleton. Lessdeformable RBCs are excluded from capillary channels resulting in lowcapillary occupancy and reduced tissue perfusion. Massive transfusion ofcells with reduced deformability may also contribute to multiple organfailure by blocking the organs' capillary beds. After transfusion,2,3-DPG is synthesized relatively quickly in vivo to ˜50% of the normallevel in as little as 7 hours and to ˜95% of the normal level in 2-3days. However, since 2,3-DPG-depleted cells do not recover their levelsimmediately, O₂-carrying capacity is compromised to the detriment ofcritically ill patients requiring immediate O₂ delivery and tissueperfusion. There are numerous reports that emphasize the importance ofRBCs with high oxygen carrying capacity in such clinical situations.

The transfusion of red blood cells (RBCs) is a life-saving therapy aimedat improving oxygenation of the tissues and vital end organs in severelyanemic patients. The majority of RBC units used for transfusion arestored at 1-6° C. for up to 42 days in an oxygen-permeablepolyvinylchloride blood bag that contains additive/preservativesolution.

Storage of frozen blood is known in the art, but such frozen blood haslimitations. For a number of years, frozen blood has been used by bloodbanks and the military for certain high-demand and rare types of blood.However, frozen blood is difficult to handle. It must be thawed thencryoprotectant must be gradually washed away which makes it impracticalfor emergency situations. Once blood is thawed, it must be used within48 hours. U.S. Pat. No. 6,413,713 to Serebrennikov is directed to amethod of storing blood at temperatures below 0° C.

U.S. Pat. No. 4,769,318 to Hamasaki et al. and U.S. Pat. No. 4,880,786to Sasakawa et al. are directed to additive solutions for bloodpreservation and activation. U.S. Pat. No. 5,624,794 to Bitensky et al.,U.S. Pat. No. 6,162,396 to Bitensky et al., and U.S. Pat. No. 5,476,764to Bitensky are directed to the storage of red blood cells underoxygen-depleted conditions. U.S. Pat. No. 5,789,151 to Bitensky et al.is directed to blood storage additive solutions. For example, Rejuvesol(available from Citra Lab LLC, Braintree, Mass.) is added to blood aftercold storage (i.e., 4° C.) just prior to transfusion or prior tofreezing (i.e., at −80° C. with glycerol) for extended storage. U.S.Pat. No. 6,447,987 to Hess et al. is directed to additive solutions forthe refrigerated storage of human red blood cells.

U.S. Pat. No. 4,837,047 to Sato et al. relates to a container forstoring blood for a long period of time to keep the quality of the bloodin good condition.

Traditional manual blood collection is performed by a trainedphlebotomist using a blood collection kit that includes, at a minimum, ablood collection bag, a phlebotomy needle, and tubing sufficient toconnect the needle to the blood collection bag containing anticoagulant.Typically, a blood collection bag further includes an anticoagulantsolution but an anticoagulant solution may alternatively be supplied ina separate bag or container connected to the blood collection bag withsuitable tubing. None of the components of current commercial systemsprovide for, or include, the reduction of oxygen.

There is a need to begin the reduction of oxygen from blood prior tostorage at the time of collection. In order to accomplish the bloodreduction within the existing infrastructure and within the time periodsas limited by current regulatory regimes, it is desirable to beginoxygen reduction as early as possible, preferably at collection beforethe temperature of the collected blood has been significantly reduced.

SUMMARY OF THE INVENTION

The present disclosure provides for, and includes, an oxygen depletiondevice for depleting oxygen from blood prior to anaerobic storagecomprising an outer receptacle substantially impermeable to oxygen, aninner collapsible blood container comprising one or more chambers thatare permeable to oxygen, and an oxygen sorbent situated within saidouter receptacle.

The present disclosure provides for, and includes, an oxygen depletiondevice for depleting oxygen from whole blood prior to anaerobic storagecomprising an outer receptacle substantially impermeable to oxygen, aninner collapsible blood container comprising one or more chambers thatare permeable to oxygen, and an oxygen sorbent situated within saidouter receptacle.

The present disclosure provides for, and includes, an oxygen depletiondevice for depleting oxygen from packed red blood cells prior toanaerobic storage comprising an outer receptacle substantiallyimpermeable to oxygen, an inner collapsible blood container comprisingone or more chambers that are permeable to oxygen, and an oxygen sorbentsituated within said outer receptacle.

The present disclosure provides for, and includes, a method to prepareblood for storage comprising providing an oxygen depletion devicecomprising an outer receptacle substantially impermeable to oxygen, aninner collapsible blood container enclosed within the outer receptacle,and an oxygen sorbent situated between the outer receptacle and innerblood compatible blood container, flowing the blood into the innercollapsible blood container of the oxygen depletion device and producingoxygen-reduced blood having less than 20% oxygen saturation.

The present disclosure provides for, and includes, a method to prepareblood for storage comprising providing an oxygen depletion devicecomprising an outer receptacle substantially impermeable to oxygen, aninner collapsible blood container enclosed within the outer receptacle,and an oxygen sorbent situated between the outer receptacle and innerblood compatible blood container, flowing the blood into the innercollapsible blood container of the oxygen depletion device and producingoxygen-reduced blood having less than 10% oxygen saturation.

The present disclosure provides for, and includes, a blood storagedevice for storing oxygen depleted blood comprising an outer receptaclesubstantially impermeable to oxygen; an inner collapsible bloodcontainer comprising a locating feature adapted to align the collapsibleblood container within the geometry of the outer receptacle; at leastone inlet comprising tubing connecting to the collapsible bloodcontainer and a bond to the outer receptacle, wherein the bond to theouter receptacle is substantially impermeable to oxygen; and an oxygensorbent situated within the outer receptacle.

The present disclosure provides for, and includes an oxygen depletiondevice 10 for depleting oxygen from blood prior to anaerobic storagecomprising an outer receptacle 101 substantially impermeable to oxygen;an oxygen indicator 206, a spacer material 110, and about 80 grams of anoxygen sorbent 103 in between an outer receptacle 101 and a 15 μm to 200μm thick silicone collapsible blood container 102.

The present disclosure provides for, and includes an oxygen depletiondevice 10 for depleting oxygen from blood prior to anaerobic storagecomprising an outer receptacle 101 substantially impermeable to oxygen;an oxygen indicator 206, a spacer material 110, and about 80 grams of anoxygen sorbent 103 in between an outer receptacle 101 and a collapsibleblood container 102 prepared from PVDF having a 0.2 μm pore size. Thepresent disclosure provides for methods to prepare blood for storagecomprising: providing an oxygen depletion device 10 and flowing bloodinto the inner collapsible blood container 102, agitating the oxygendepletion device 10 for up to 3 hours, producing oxygen-reduced bloodhaving less than 20% oxygen saturation, and transferring theoxygen-reduced blood to a blood storage device 20. The method furtherprovides for the production of oxygen-reduced blood having less than 20%oxygen saturation in less than 8 hours after collection from a donor. Ina further embodiment, the agitating is nutating.

The present disclosure provides for, and includes a method of reducingoxygen from whole blood, or a component thereof, comprising placing thewhole blood, or a component thereof in a device 20 comprising a sorbent207 that has an absorption rate of at least 1.86 cubic centimeters pergram sorbent per hour (cc·g⁻¹·hr⁻¹), incubating the blood filled device20 for up to four hours at ambient temperature while agitating at leastonce per second by translation of at least 3 cm; transferring the bloodfilled device 20 to storage at 4 to 6° C. In a further aspect, the bloodfilled device 20 is stored at 4 to 6° C. for up to 42 days.

BRIEF DESCRIPTION OF THE DRAWINGS

Some aspects of the disclosure are herein described, by way of exampleonly, with reference to the accompanying drawings. With specificreference now to the drawings in detail, it is stressed that theparticulars shown are by way of example and are for purposes ofillustrative discussion of embodiments of the disclosure. In thisregard, the description, taken with the drawings, makes apparent tothose skilled in the art how aspects of the disclosure may be practiced.

FIGS. 1A-C illustrate an exemplary embodiment of an oxygen depletiondevice according to the present disclosure having two compartmentsarranged side by side.

FIGS. 2A and 2B illustrate an exemplary embodiment of an oxygendepletion device according to the present disclosure having threecompartments arranged side by side.

FIGS. 3A and 3B illustrate an exemplary embodiment of an anaerobicstorage bag according to the present disclosure.

FIGS. 4A and 4B illustrate an exemplary embodiment of an oxygenreduction disposable storage system having a blood depletion devicehaving two or three compartments, respectively, and an anaerobic storagebag according to the present disclosure.

FIG. 5 is a graph of sO₂ reduction in exemplary oxygen depletion devicesaccording to the methods of the present disclosure.

FIGS. 6A and 6B illustrates exemplary embodiments of an anaerobicstorage bag according to the present disclosure.

FIG. 7 illustrates an exemplary embodiment of tie layers 105 joiningmembranes 113 and 114 in a two-step process according to the presentdisclosure.

FIGS. 8A and 8B illustrate an exemplary embodiment of a spacer 110comprising inner mesh 117 coextruded with binder mesh 118 and joined toa membrane 113 (114) according to the present disclosure.

FIGS. 9A and 9B illustrate an exemplary embodiment of an anaerobicstorage bag having a tie layer 105 joining membranes 113 and 114 (9A)and tie layers 105 applied to membranes 113 and 114 providing a seal 108wherein the tie layers 105 extend beyond the seal 108 by a distance 109(9B) according to the present disclosure.

FIGS. 10A to 10D illustrate exemplary embodiments of a tie layer 105having geometric features 121 according to the present disclosure andfurther comprising a mixing structure 109 as shown in 10C and 10D.

FIG. 11 illustrates an exemplary embodiment of a collapsible bloodcontainer having a spacer 110, a tie layer 105, and a geometric feature121 according to the present disclosure.

FIG. 12 is a graph of sO₂ reduction in an exemplary oxygen depletiondevice according to the methods of the present disclosure.

FIG. 13 is a graph of sO₂ reduction in an exemplary inner collapsibleblood container 102 with various blood volumes, according to the methodsof the present disclosure.

FIG. 14 is a graph of sO₂ reduction in an exemplary oxygen depletiondevice according to the methods of the present disclosure.

FIG. 15 is a graph of sO₂ reduction in an exemplary oxygen depletiondevice having different surface areas according to the methods of thepresent disclosure.

FIG. 16 is a graph showing the effect of spacer 110 on sO₂ reduction inan exemplary oxygen depletion device according to the presentdisclosure.

Corresponding reference characters indicate corresponding partsthroughout the several views. The examples set out herein illustrateseveral embodiments of the invention but should not be construed aslimiting the scope of the invention in any manner.

In light of current technology, there is a need to improve the qualityof blood and blood components such as red blood cells that are to bestored and to extend the storage life of such blood and blood componentsin advance of transfusion to help minimize morbidity associated withtransfusions. In order to conform with regulatory requirements and toensure reliability, the preparation and processing of the red bloodcells must be completed within a limited time period. Further, theprocess of preparing reduced oxygen blood and blood components must notintroduce lesions, including but not limited to, hemolysis of the blood.Finally, there is a need for methods and devices that are compatiblewith existing anticoagulant and additive solutions to yield improvedquality blood and blood components.

DETAILED DESCRIPTION

To address such needs and others, the present disclosure includes andprovides devices and methodology for the preservation of blood and bloodcomponents in which the preparation of oxygen reduced blood and bloodcomponents is initiated at the donor collection stage.

Before explaining at least one aspect of the disclosure in detail, it isto be understood that the disclosure is not necessarily limited in itsapplication to the details set forth in the following description orexemplified by the Examples. The disclosure is capable of other aspectsor of being practiced or carried out in various ways.

As used herein, the term “bag” refers to collapsible containers preparedfrom a flexible material and includes pouches, tubes, and gusset bags.As used herein, and included in the present disclosure, the termincludes folded bags having one, two, three, or more folds and which aresealed or bonded on one, two, three, or more sides. Bags may be preparedusing a variety of techniques known in the art including bonding ofsheets of one or more materials. Methods of bonding materials to formbags are known in the art. Also included and provided for in the presentdisclosure are containers prepared by injection and blow molding.Methods to prepare blow molded and injection molded containers are knownin the art. Preferred types of blow molded or injection moldedcontainers are flexible containers that can be reduced in size forefficient packing and shipping while being capable of expanding toaccommodate blood or blood components for reduction of oxygen. They alsomay be designed to conform to the volume of the blood until they arefully expanded. As used throughout the present disclosure, the bags area form of collapsible container and the two terms are usedinterchangeably throughout the present disclosure.

As used herein, the term “collapsible container” includes bags,containers, enclosures, envelopes, pouches, pockets, receptacles, andother devices that can contain and retain a liquid or fluid. In certainaspects, the collapsible container may be manufactured by conventionalmeans such as injection molding or insert molding. In other aspects, thecollapsible container may be prepared from sheets of polymer materialsthat are bonded together using methods known in the art to preparecontainers capable of holding a volume. Such collapsible containers arewell known in the art. See, for example, U.S. Pat. No. 3,942,529 issuedto Waage; U.S. Pat. No. 4,131,200 issued to Rinfret; and U.S. Pat. No.5,382,526 issued to Gajewski et al. Suitable methods for bonding polymermaterials to prepare collapsible containers according to the presentdisclosure include heat welding, ultrasonic welding, radio frequency(RF) welding, and solvent welding. In certain aspects, multiple bondingmethods may be used to prepare collapsible containers according to thepresent disclosure. Collapsible containers according to the presentdisclosure include enclosures having one or more pleats, folds,diaphragms, bubbles, and gussets. Methods for preparing collapsiblecontainers are known in the art. See, for example, U.S. Pat. No.3,361,041 issued to Grob; U.S. Pat. No. 4,731,978 issued to Martensson;U.S. Pat. No. 4,998,990 issued to Richter et al.; and U.S. Pat. No.4,262,581 issued to Ferrell. Also included and provided for in thepresent disclosure are containers having combinations of both flexibleand inflexible parts, wherein the flexible parts allow for the expansionof the volume through, for example, pleats, folds or gussets and othersimilar geometric features in the packaging shape, whereas theinflexible parts may provide rigidity and geometry definition to thecontainer. Methods and designs for preparing collapsible containershaving both flexible and inflexible parts are known in the art, such asdescribed by Randall in U.S. Pat. No. 6,164,821 and by LaFleur in U.S.Pat. No. 5,328,268.

As used herein the term “about” refers to ±10%.

The terms “comprises,” “comprising,” “includes,” “including,” “having,”and their conjugates mean “including but not limited to.”

The term “consisting of” means “including and limited to.”

The term “consisting essentially of” means that the composition, methodor structure may include additional ingredients, steps and/or parts, butonly if the additional ingredients, steps and/or parts do not materiallyalter the basic and novel characteristics of the claimed composition,method or structure.

As used herein, the singular forms “a,” “an,” and “the” include pluralreferences unless the context clearly dictates otherwise. For example,the term “a compound” or “at least one compound” may include a pluralityof compounds, including mixtures thereof.

Throughout this application, various embodiments of this disclosure maybe presented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of thedisclosure. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible subranges as well asindividual numerical values within that range. For example, descriptionof a range such as “from 1 to 6” should be considered to havespecifically disclosed subranges such as “from 1 to 3,” “from 1 to 4,”“from 1 to 5,” “from 2 to 4,” “from 2 to 6,” “from 3 to 6,” etc., aswell as individual numbers within that range, for example, 1, 2, 3, 4,5, and 6. This applies regardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to includeany cited numeral (fractional or integral) within the indicated range.The phrases “ranging/ranges between” a first indicate number and asecond indicate number and “ranging/ranges from” a first indicate number“to” a second indicate number are used herein interchangeably and aremeant to include the first and second indicated numbers and all thefractional and integral numerals therebetween.

As used herein the term “method” refers to manners, means, techniques,and procedures for accomplishing a given task including, but not limitedto, those manners, means, techniques, and procedures either known to orreadily developed from known manners, means, techniques, and proceduresby practitioners of the chemical, pharmacological, biological,biochemical, and medical arts.

The present disclosure provides for, and includes, an oxygen depletiondevice 10 for depleting oxygen from blood comprising an outer receptacle101 substantially impermeable to oxygen, inner collapsible bloodcontainer 102 that is permeable to oxygen, and an oxygen sorbent 103situated within outer receptacle 101.

The present disclosure also provides for, and includes, oxygen depletiondevices 10 configured to be a blood collection and oxygen depletiondevice 10. Oxygen depletion devices configured to collect and reduceblood oxygen differ from the oxygen depletion device 10 as describedthroughout this specification in that a blood collection and oxygendepletion device 10 further includes an anticoagulant to preventcoagulation of the whole blood during the collection process. In certainaspects, the anticoagulant solution of a blood collection and oxygendepletion device 10 is provided in the blood collection and oxygendepletion device 10. Accordingly, included anticoagulant solutions arealso oxygen depleted anticoagulant solutions. In the alternative,anticoagulant solutions may be included separately, either as oxygendepleted solutions or solutions having oxygen. A blood collection andoxygen depletion device 10 is intended to be used with whole bloodcollected from a donor. As used throughout the present disclosure, theoxygen and depletion device 10 includes and provides for bloodcollection and oxygen depletion device 10. The two terms can be, andare, used interchangeably.

As used herein, the outer receptacles are prepared from materials thatare substantially impermeable to oxygen and optionally impermeable tocarbon dioxide. In certain aspects, an outer receptacle 101 is preparedfrom flexible film materials. In other aspects, an outer receptacle 101is prepared from a stiff, or inflexible film material.

The present disclosure provides for, and includes, an outer receptacle101 substantially impermeable to oxygen. As used herein, an outerreceptacle 101 that is substantially impermeable to oxygen issufficiently impermeable to oxygen to allow no more than 10 cc of oxygeninside the receptacle over a period of 3 months, and more preferably nomore than 5 cc of oxygen over 6 months. As used herein, the termsubstantially impermeable to oxygen (SIO) refers to materials andcompositions that provide a barrier to the passage of oxygen from oneside of the barrier to the other, sufficient to prevent significantincreases in the partial pressure of oxygen.

It is notable that few materials provide complete impermeability andthat even the high impermeability of materials can be compromised whenjoining, welding, folding, and otherwise assembling an outer receptacle101. As will be discussed below, oxygen depletion device 10 may furtherincorporate one or more inlets/outlets 30 comprising a tube 301 and abond 302 to the outer receptacle 101 (or outer receptacle 201 describedbelow). The outer receptacle 101 must also be designed to accommodatechanges in volume of the inner collapsible blood container 102.Accordingly, special care is taken to incorporate specific designelements and manufacturing methods to ensure the integrity of theimpermeable barrier.

The present disclosure also provides for, and includes, an outerreceptacle 101 that is substantially impermeable to oxygen having apermeability to oxygen of less than about 1.0 cc of oxygen per squaremeter per day. In certain aspects, a film suitable for use in thepreparation of an outer receptacle and other elements of the presentdisclosure are materials characterized by a Barrer value of less thanabout 0.140 Barrer.

Materials and methods to prepare an outer receptacle 101 are known inthe art. See, for example, U.S. Pat. No. 7,041,800 issued to Gawryl etal., U.S. Pat. No. 6,007,529 issued to Gustafsson et al., and U.S.Patent Application Publication No. 2013/0327677 by McDorman, each ofwhich are hereby incorporated by reference in their entireties.Impermeable materials are routinely used in the art and any suitablematerial can be used. In the case of molded polymers, additives areroutinely added to enhance the oxygen (and CO₂) barrier properties. See,for example, U.S. Pat. No. 4,837,047 issued to Sato et al. For example,U.S. Pat. No. 7,431,995 issued to Smith et al. describes an oxygen- andcarbon dioxide-impermeable receptacle composed of layers of ethylenevinyl alcohol copolymer and modified ethylene vinyl acetate copolymer,impermeable to oxygen and carbon dioxide ingress. In another aspect, theouter receptacle 101 is impermeable to oxygen and carbon dioxide.

In certain aspects, films that are substantially impermeable to oxygenmay be laminated films. In an aspect, a laminated film that issubstantially impermeable to oxygen is a laminated foil film. Filmmaterials can be polymers or foil materials or multilayer constructionsthat are combinations of foils and polymers. In an aspect, a laminatedfilm may be a polyester membrane laminated with aluminum. An example ofsuitable aluminum laminated film, also known as a laminated foil, thatis substantially impermeable to oxygen is known in the art. For example,U.S. Pat. No. 4,798,728 to Sugisawa discloses aluminum laminated foilsof nylon, polyethylene, polyester, polypropylene, and vinylidenechloride. Other laminated films are known in the art. For example, U.S.Pat. No. 7,713,614 to Chow et al. discloses multilayer containerscomprising an ethylene-vinyl alcohol copolymer (EVOH) resin that issubstantially impermeable to oxygen. In an aspect, an outer receptacle101 may be a barrier bag constructed by sealing three or four sides bymeans of heat sealing. The bag is constructed of a multilayerconstruction that includes materials that provide enhancement to O₂ andCO₂ barrier properties. The bag is constructed of a multilayerconstruction that includes materials that provide enhancement to O₂ andCO₂ barrier properties. Such materials include the Rollprint Clearfoil®V2 film, having an oxygen transmission rate of 0.01 cc/100 in²/24 hrs.,Rollprint Clearfoil® X film, having an oxygen transmission rate of 0.004cc/100 in²/24 hrs. and Clearfoil® Z film having an oxygen transmissionrate of 0.0008 cc/100 in²/24 hrs. (Rollprint Packaging Products,Addison, Ill.). Other manufacturers make similar products with similaroxygen transmission rates, such as Renolit Solmed Wrapflex® films(American Renolit Corp., City of Commerce, Calif.). An example ofsuitable aluminum laminated film, also known as a laminated foil, thatis substantially impermeable to oxygen is obtainable from ProtectivePackaging Corp. (Carrollton, Tex.).

Another approach applicable to the preparation of SIO materials includesmultilayer graphitic films made by gentle chemical reduction of grapheneoxide laminates with hydroiodic and ascorbic acids. See Su et al.,“Impermeable barrier films and protective coatings based on reducedgraphene oxide,” Nature Communications 5, Article number: 4843 (2014),hereby incorporated by reference in its entirety. Nanoparticles toenhance oxygen barrier properties are also known in the art, forexample, the multilayer barrier stack films provided by Tera-Barrier(Tera-Barrier Films Pte, Ltd, The Aries, Singapore) and described byRick Lingle in Packaging Digest Magazine on Aug. 12, 2014.

In aspects according to the present disclosure, an outer receptacle 101may be prepared from a gas impermeable plastic. In an embodiment, thegas impermeable plastic may be a laminate. In certain embodiments, thelaminate may be a transparent barrier film, for example, a nylonpolymer. In embodiment, the laminate may be a polyester film. In anembodiment, the laminate may be Mylar®. In certain embodiments, thelaminate may be a metalized film. In an embodiment, the metalized filmmay be coated with aluminum. In another embodiment, the coating may bealuminum oxide. In another embodiment, the coating may be an ethylenevinyl alcohol copolymer (EVOH) laminated between layers of low densitypolyethylene (LDPE).

An outer receptacle 101 of the present disclosure may be formed of oneor more parts prepared from a gas impermeable material including aplastic or other durable lightweight material. In some embodiments, anenclosure may be formed of more than one material. In an embodiment, anouter receptacle 101 may be formed of a material and coated with a gasimpermeable material to prepare a gas impermeable enclosure. In anembodiment, a rigid or flexible outer receptacle 101 may be preparedfrom a plastic that may be injection molded. In embodiments according tothe instant disclosure, the plastic may be selected from polystyrene,polyvinyl chloride, or nylon. In an embodiment, outer receptacle 101materials may be selected from the group consisting of polyester (PES),polyethylene terephthalate (PET), polyethylene (PE), high-densitypolyethylene (HDPE), polyvinyl chloride (PVC), polyvinylidene chloride(PVDC), low-density polyethylene (LDPE), polypropylene (PP), polystyrene(PS), high impact polystyrene (HIPS), polyamides (PA) (e.g., nylon),acrylonitrile butadiene styrene (ABS), polycarbonate (PC),polycarbonate/acrylonitrile butadiene styrene (PC/ABS), polyurethanes(PU), melamine formaldehyde (MF), plastarch material, phenolics (PF),polyetheretherketone (PEEK), polyetherimide (PEI) (Ultem), polylacticacid (PLA), polymethyl methacrylate (PMMA), polytetrafluoroethylene(PTFE), urea-formaldehyde, and ethylene vinyl alcohol copolymer (EVOH).In certain embodiments, the outer receptacle 101 may be polyethylene. Insome embodiments, the polyethylene outer receptacle 101 may comprise oneor more polyethylene components that are welded together. In certainaspects, the outer receptacle is comprised of a multilayer film having apolyethylene outer layer, a polyester inner layer, and an aluminum oxidebarrier layer dispersed between the inner and outer layers, for example,the Clearfoil® Z film having an oxygen transmission rate of 0.0008cc/100 in²/24 hrs. (Rollprint Packaging Products, Addison, Ill.).

The present disclosure provides for, and includes, the preparation ofouter receptacles 101 from a film and inner collapsible blood container102 from a membrane. As used herein, membranes generally refer tomaterials used to prepare an inner collapsible blood container 102 andfilms are used to refer to materials used to prepare outer receptacle101. While it is understood that certain materials may be referred bythe manufacturer as a “membrane” or may be generally known as a“membrane”, for clarity, unless otherwise indicated a film is consideredsubstantially impermeable. A membrane comprises one or more layers ofmaterials in the form of a sheet that allows one or more substances topass through from one side of the sheet to the other side of the sheet.As used herein, membranes may also be prepared as tubes suitable forconnecting together components of oxygen depletion devices 10, bloodcollection kits, or connecting together elements of blood collectiondevices, additive solution bags, leukocyte reduction filters, andanaerobic storage bags. As used throughout, it is understood that amembrane of the present disclosure may be formed as a sheet or a tubedepending on the application. Also as previously provided, films toprepare outer receptacles 101 are substantially impermeable to oxygenwhile an inner collapsible blood container 102 is permeable to oxygen.As used herein, films may also be prepared as tubes suitable forconnecting together components of oxygen depletion devices 10, bloodcollection kits, or connecting together elements of blood collectiondevices, additive solution bags, leukocyte reduction filters, andanaerobic storage bags. As used herein the outer receptacles 101 containall embodiments of 102 as further described herein.

As used herein, an inner collapsible blood container 102 is permeable tooxygen. In certain aspects, an inner collapsible blood container 102 ispermeable to oxygen and carbon dioxide. In other aspects, an innercollapsible blood container 102 is impermeable to oxygen and permeableto carbon dioxide.

The present disclosure provides for and includes the preparation ofouter receptacles 101 using heat sealing, blow molding, and injectionmolding techniques. Suitable materials for preparing outer receptacles101 using heat sealing, blow molding, and injection molding include PET,standard and multilayer, polypropylene, polyethylene, polycarbonate,ABS, and other polymers known to those skilled in the art. Methods toprepare blow molded and injection molded outer receptacles 101 are knownin the art, for example, a multilayer structure comprised of a barrierlayer of ethylvinyl alcohol (EVOH) or ethylvinylacetate (EVA) situatedbetween two layers of polypropylene (PP) and offered by Kortec (Kortec,Inc., Rowley, Mass.) and also as described in U.S. Pat. No. 5,906,285issued to Slat. Additives that strengthen the oxygen and CO₂ barrierproperties of the polymers prior to molding or during their formulationor during setup are known in the art. One example is multilayer polymerco-injection resulting in a multilayer PET. Such a barrier resin istypically incorporated at the preform stage as an inner layer with PETon both sides, making PET the liquid contact layer as well as theoutside layer. As provided below, suitable blow molded or injectionmolded outer receptacles 101 are impermeable to oxygen. In certainaspects, suitable heat sealed, blow molded, or injection molded outerreceptacles 101 are substantially impermeable to both oxygen and carbondioxide.

The present disclosure provides for, and includes, two types ofmaterials for the preparation of either permeable membranes orsubstantially impermeable films. In an aspect, permeable membranesaccording to the present disclosure provide for the passage ofsubstances through the material, specifically but not necessarilyexclusively, oxygen. In certain aspects, membranes are selected topermit the passage of oxygen and carbon dioxide while preventing thepassage of water, proteins, salts (e.g., plasma components) and cells(e.g., red blood cells, white blood cells, and platelets). The rate ofpassage through a material depends on one or more properties includingparticle size, phase of material (liquid vs. gas), hydrophilicity,hydrophobicity, or solubility. The rate of passage, or flux, through amaterial also depends on the presence or absence of a driving force suchas a difference in pressure (or partial pressure), differences intemperature, or differences in concentration between one side of themembrane and the other. The flux through a membrane is known as themembrane permeation flux. The membrane permeation flux of substancesthrough a membrane is inversely proportional to the thickness of themembrane.

Membrane permeation flux, for a gas, is defined as the volume flowingthrough the membrane per unit area per unit time. The SI unit used ism³/m²·s. For gases and vapors, the volume is strongly dependent onpressure and temperature. Accordingly, permeation fluxes for gases areoften given in terms of standard temperature and pressure (STP) which isdefined as 0° C. and 1 atmosphere (1.0013 bar) (e.g., 273° K. and 760torr). As noted above, the rate of passage depends on a driving force ordifference between the two sides of the membrane, and this dependence isincorporated in the permeability coefficient, P, or simply thepermeability.

Permeability (P) is defined as the permeability flux per unit of drivingforce per unit of membrane thickness. The SI unit for the permeabilitycoefficient P is provided in Table 1. A common unit for gas separation,as in the present disclosure, is the Barrer and is also presented inTable 1. The term cm³ gas (STP)/cm²s refers to the volumetrictrans-membrane flux of the diffusing species in terms of standardconditions of 0° C. and 1 atmosphere pressure, the term cm refers to themembrane thickness, and cm-Hg refers to the trans-membrane partialpressure driving force for the diffusing species. Permeability must beexperimentally determined.

TABLE 1 Permeability Units Units of Permeability “Volumetric”permeability ${1\mspace{14mu}{Barrer}} = \frac{\begin{matrix}{{10^{- 10} \cdot {cm}^{3}}{{{gas}({STP})} \cdot}} \\( {{cm}\mspace{14mu}{membrane}\mspace{14mu}{thickness}} )\end{matrix}}{( {{cm}^{2}\mspace{14mu}{membrane}\mspace{14mu}{area}} ) \cdot s \cdot ( {{cmHg}\mspace{14mu}{pressure}} )}$“Molar” permeability${\frac{mol}{m \cdot {Pa} \cdot s}( {{SI}\mspace{14mu}{units}} )} = \frac{\begin{matrix}{( {{mol}_{i}\mspace{14mu}{permeating}} ) \cdot} \\( {m\mspace{14mu}{membrane}\mspace{14mu}{thickness}} )\end{matrix}}{( {m^{2}\mspace{14mu}{membrane}\mspace{14mu}{area}} ) \cdot s \cdot ( {{Pa}\mspace{14mu}{pressure}} )}$

Membranes suitable for the methods and devices according to the presentdisclosure include dense membranes, porous membranes, asymmetricmembranes, and composite membranes. In certain aspects, suitablemembranes may be multilayered membranes. In other aspects, suitablemembranes are prepared from inorganic materials. Dense membranes aremembranes prepared from solid materials that do not have pores or voids.Materials permeate dense membranes by processes of solution anddiffusion. Examples of dense membranes include silicone membranes(polydimethyl siloxane, or PDMS). Also included and provided for in thepresent disclosure are porous membranes that have pores of a particularrange of sizes that separate on the basis of size exclusion. Examples ofporous membranes suitable for use according to the present disclosureinclude PVDF and polysulfone membranes.

Included and provided for by the present disclosure are compositemembranes that are made of more than one material, often as laminates,wherein a dense material is applied to a porous support layer. Examplesof composite membranes suitable for use according to the presentdisclosure are EMD Millipore's GVHP hydrophobic PVDF having 1.0 μm or0.22 μm pore sizes.

TABLE 2 Permeability of Fluoropolymers (100 μm thick; 23° C.) SiliconePTFE PFA FEP ETFE CTFE ECTFE PVDF PVF THV Water vapor 36000 5 8 1 2 1 22 7 1.73 (g/m² · d · bar) Oxygen 500 1500 n/a 2900 350 60 100 20 12 696(cm³/m² · d · bar) Nitrogen 280 500 n/a 1200 120 10 40 30 1 217 (cm³/m²· d · bar) CO₂ 2700 15000 7000 4700 1300 150 400 100 60 2060 (cm³/m² · d· bar) From Kunststoffe “Fluorocarbon films - Present situation andFuture Outlook” available at kynar.com

The present disclosure provides for, and includes, inner collapsibleblood containers 102 prepared from membranes 113 that are characterizedprimarily by their permeability to oxygen. Unless indicated otherwise, a“substantially impermeable membrane” refers to membranes that aresubstantially impermeable to oxygen. However, in certain devices andmethods, the membranes may be further characterized by the permeabilityor impermeability to carbon dioxide. For certain applications, themembrane material is substantially impermeable to oxygen and provides abarrier to the introduction of oxygen to the blood, blood component, ora blood collection kit comprised of multiple components. Suchsubstantially impermeable membranes are generally used to prepare outerreceptacles of the present disclosure. Suitable substantiallyimpermeable membranes may also be used to prepare tubing for connectivecomponents of the devices and kits. Substantially impermeable membranesmay comprise a monolayer or be laminated sheets or tubes having two ormore layers.

The present disclosure also provides for, and includes, membranes 113that are substantially permeable to oxygen. Membranes 113 that aresubstantially permeable to oxygen are used in the present disclosure forthe preparation of inner collapsible blood containers 102. In certainaspects, the membranes 113 that are permeable to oxygen are alsobiocompatible membranes, approved and suitable for extended contact withblood that is to be transfused into a patient. Like substantiallyimpermeable membranes, substantially permeable membranes 113 maycomprise a monolayer or may comprise a laminated structure having two ormore layers.

In an aspect, oxygen permeable membranes 113 having a permeability tooxygen of greater than about 2.5×10⁻⁹ cm³ O₂ (STP)/((cm² s)*(cm Hgcm⁻¹)) is used for the preparation of a collapsible blood container 102.In another aspect, oxygen permeable membranes 113 having a permeabilityto oxygen greater than about 5.0×10⁻⁹ cm³ O₂ (STP)/((cm² s)*(cm Hgcm⁻¹)) is used for the preparation of a collapsible blood container 102.In yet another aspect, oxygen permeable membranes 113 have apermeability to oxygen of greater than about 1.0×10⁻⁸ cm³ O₂ (STP)/((cm²s)*(cm Hg cm⁻¹)). In certain aspects, oxygen permeable membranes 113suitable for use in the preparation of a collapsible blood container 102are characterized by a Barrer value of greater than about 25. In otheraspects, oxygen permeable membranes 113 suitable for use in thepreparation of a collapsible blood container 102 are characterized by aBarrer value of greater than about 50. In certain other aspects, oxygenpermeable membranes 113 suitable for use in the preparation of acollapsible blood container 102 are characterized by a Barrer value ofgreater than about 100.

In an aspect, a membrane 113 that is substantially permeable to oxygencan be dense membranes prepared from non-porous materials. Examples ofsuitable materials that are capable of high oxygen permeability ratesinclude silicones, polyolefins, epoxies, and polyesters. In anotheraspect, membranes that are substantially permeable to oxygen can beporous membranes prepared from organic polymers. A membrane 113 that issubstantially permeable to oxygen may be prepared from a materialselected from the group consisting of PVDF rendered hydrophobic, nylon,cellulose esters, polysulfone, polyethersulfone, polypropylene renderedhydrophobic, and polyacrylonitrile.

The present disclosure provides for, and includes, preparing membranes113 that are substantially permeable to oxygen, not only by selectingthe material, but also by selecting and controlling the thickness. Asprovided above, permeability is proportional to the thickness of themembrane. Accordingly, improved permeability may be achieved bydecreasing the thickness of the membrane. In certain aspects, theminimum thickness is determined by its strength and resistance topuncture and tearing.

The present disclosure also provides for, and includes, membranes 113that are substantially permeable to oxygen that are prepared using blowmolding and injection molding techniques. Suitable materials forpreparing inner collapsible blood containers 102 using blow molding andinjection molding include silicone materials such as Bluestar 4350, 50durometer, Silbione grade liquid silicone rubber and Shin-EtsuKEG-2000-40A/B Liquid Silicone. The silicone durometer choice iscarefully chosen for collapsibility and permeability, followed by awell-controlled wall thickness. Thinner materials will have a higherpermeability. Methods to prepare blow molded and injection moldedcollapsible blood containers 102 are known in the art, for example, U.S.Pat. No. 4,398,642 issued to Okudaira et al.; U.S. Pat. No. 7,666,486issued to Sato et al.; U.S. Pat. No. 8,864,735 issued to Sano et al.;and U.S. Patent Application Publication No. 2012/0146266 by Oda et al.In an aspect, a blow molded collapsible blood container 102 can beprepared using LDPE used in the manufacture of collapsible watercontainers. As provided below, suitable blow molded or injection moldedcollapsible blood containers 102 have a permeability to oxygen of atleast about 25 Barrer.

In an aspect according to the present disclosure, the collapsible bloodcontainer 102 can be manufactured from microporous membrane 113 byvarious sealing methods such as heat sealing, thermal staking, andadhesive bonding. In one aspect according to the present disclosure, apair of PVDF microporous membranes are bonded together around theperiphery with a section of PVC inlet tubing in place in the seam usingan adhesive such as Loctite 4011 in conjunction with an adhesive primersuch as Loctite 770. In another aspect according to the presentdisclosure, a collapsible blood container can be manufactured from apair of microporous membranes by heat sealing the 4 edges of the pair ofmembranes together with a section of multilayer tubing sealed into theseam to provide for fluid connectivity.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is prepared from more than one type of membrane 113.In an aspect, a collapsible blood container 102 comprises a firstmembrane 113 and a second membrane 114 suitably bonded to prepare acontainer. As used herein, a membrane 114 generally refers to a membranethat is identical to membrane 113. That is, a collapsible bloodcontainer 102 is generally made of two joined membranes 113. The presentdisclosure provides for, and includes, a collapsible blood container 102that is prepared from a membrane 113 and a membrane 114 comprising adifferent material. As shown in FIG. 1C, a collapsible blood container102 is shown to be prepared with a membrane 113 and a membrane 114.Unless indicated otherwise, it is understood that a membrane 113 and amembrane 114 may be exchanged. In another aspect, a collapsible bloodcontainer 102 comprises a membrane 113 combined with a second membrane114 that has a permeability of less than about 30% of the permeabilityof first membrane 113. In certain aspects, a second membrane 114comprises a membrane that is relatively impermeable or insufficientlypermeable to provide sufficient deoxygenation on its own, but can becombined with a suitable membrane 113. In certain aspects, the secondmembrane 114 is relatively impermeable. In further aspects, the secondmembrane 114 comprises a molded membrane that incorporates ridges,baffles, or other structures to facilitate mixing. In an aspect, thesecond membrane 114 may comprise a rigid structure joined to an oxygenpermeable membrane 113. In aspects according to the present disclosure,the second membrane 114 is heat sealed to membrane 113.

In certain aspects, the inner collapsible blood container 102 containsflow baffles located internal or external to the blood contact area thatprovide an increase in the turbulence inside the collapsible bloodcontainer 102 when agitated. In an aspect, baffles are located 1 to 2inches from each other and comprise 10 to 45% of the inner collapsibleblood container 102 area.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is substantially permeable to oxygen and is amicroporous membrane prepared from polyvinylidene fluoride, orpolyvinylidene difluoride (PVDF). In certain aspects, the PVDF membraneis a hydrophobic microporous membrane that is substantially permeable tooxygen.

In aspects according to the present disclosure, the microporous PVDFmembrane comprises pores having a range of between 0.01 μm and 2.0 μm.In other aspects, the microporous PVDF membrane 113 comprises poreshaving a range of between 0.01 μm and 1.0 μm. In some aspects, amicroporous PVDF membrane 113 has a pore size of between 0.03 μm and 1.0μm in diameter. In other aspects, a microporous PVDF membrane 113 has apore size of between 0.03 μm and 0.45 μm in diameter.

In aspects according to the present disclosure, the void fraction of aPVDF membrane 113 used to prepare a collapsible blood container 102 isbetween 20 and 80%. In another aspect, the void fraction of a PVDFmembrane 113 used to prepare a collapsible blood container 102 isbetween 35 and 50%.

In certain aspects, the permeability of PVDF membranes having microporesgreater than about 1.0 μm may allow fluid to permeate through themembrane, compromising both the fluid containment as well as the oxygenand carbon dioxide permeability. To overcome this permeability at highpore sizes, so called “super-hydrophobic” membranes can be employedwherein the contact angle is greater than 150°. As used herein and knownin the art, the contact angle quantifies the wettability of a solidsurface and is theoretically described by Young's equation. In certainaspects according the present disclosure, the use of non-hydrophobicPVDF materials is not recommended as the surface tension of the materialis lower and allows for fluid to seep through the pores even at theranges stated above.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a PVDF permeable membrane 113having a pore size of between 0.1 and 0.8 μm in diameter. In otheraspects, micropores of porous PVDF membranes may be from 0.22 to 0.8 μmin diameter. In an aspect, the micropores of porous PVDF membranes arefrom 0.2 to 1.0 μm. In another aspect, the micropores of porous PVDFmembranes may be greater than 0.1 and less than 1.0 μm. In a furtheraspect, the micropore of the porous PVDF membrane ranges from about 0.05to about 1.0 μm. In some aspects, the micropores of porous PVDFmembranes may be greater than 0.3 or 0.4 μm. In other aspects, themicropores of porous PVDF membranes may be greater than 0.5 or 0.6 μm.

In aspects according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PVDF membrane 113 having a micropore size of less than 1.0 μm. Inanother aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PVDF membrane 113 having a micropore size of less than 0.8 μm. Incertain aspects according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PVDF membrane 113 having a micropore size of less than 0.65 μm. Inanother aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PVDF membrane 113 having a micropore size of less than 0.45 μm.

In an aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PVDF membrane 113 having a micropore size of 0.1 μm. In anotheraspect, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a PVDF membrane 113 having a microporesize of 0.22 μm. In another aspect, an oxygen depletion device 10comprises an inner collapsible blood container 102 comprising a PVDFmembrane 113 having a micropore size of 0.20 μm. In a further aspectaccording to the present disclosure, an oxygen depletion device 10comprises an inner collapsible blood container 102 comprising a PVDFmembrane 113 having a micropore size of 0.45 μm. In yet a furtheraspect, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a PVDF membrane 113 having a microporesize of 0.65 μm. In another aspect according to the present disclosure,an oxygen depletion device 10 comprises an inner collapsible bloodcontainer 102 comprising a PVDF membrane 113 having a micropore size of0.8 μm.

In aspects according to the present disclosure, the PVDF membrane may beless than 250 μm thick. In certain aspects, the membrane is greater than10 μm thick. In some aspects, the PVDF membrane may be between 10 and250 μm thick. In other aspects, the PVDF membrane may be between 10 and125 μm thick or between 25 and 150 μm thick. In an aspect, the PVDFmembrane may be between 50 and 125 μm thick, 75 and 125 μm thick, 50 and150 μm thick, 75 and 150 μm thick, 100 and 125 μm thick, 150 and 250 μmthick, or between 25 and 150 μm thick. In an aspect, the membrane 113 ofinner collapsible blood container 102 is about 20 μm thick. In anotheraspect, the membrane 113 of inner collapsible blood container 102 isabout 30 μm thick. In yet another aspect, the membrane 113 of innercollapsible blood container 102 is about 50 μm thick. In a furtheraspect, the membrane 113 of inner collapsible blood container 102 isabout 76 μm thick. In an aspect, the membrane 113 of inner collapsibleblood container 102 is about 120 μm thick.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a PVDF permeable membrane 113 thatis between 100 and 125 μm thick. In certain aspects according to thepresent disclosure, the collapsible blood container 102 is prepared froma PVDF permeable membrane 113 having a pore size of between 0.1 μm and0.8 μm in diameter and that is between 100 and 125 μm thick. In certainaspects according to the present disclosure, the collapsible bloodcontainer 102 is prepared from a PVDF permeable membrane 113 having apore size of between 0.1 μm and 0.8 μm in diameter and that is between50 and 150 μm thick.

Examples of suitable PVDF membranes for the preparation of innercollapsible blood containers that are permeable to oxygen according tothe present disclosure include VVSP 115 μm thick/0.1 μm pore; GVSP 115μm thick/0.22 μm pore; HVSP 115 μm thick/0.45 μm pore; DVSP 115 μmthick/0.65 μm pore; BVSP 115 μm thick/1.0 μm pore; VVHP 107 μm thick/0.1μm pore; GVHP 125 μm thick/0.22 μm pore; HVHP 115 μm thick/0.45 μm pore;or DVHP 115 μm thick/0.65 μm pore.

Suitable PVDF membranes include commercially available membranes.Non-limiting examples of PVDF membranes are available from MilliporeCorporation, Bedford, Mass. In an aspect, the PVDF membrane may beobtained from Millipore Corporation, Bedford, Mass. An example of such aPVDF membrane is the VVSP, GVSP, HVSP, DVSP, BVSP, VVHP, GVHP, HVHP, orDVHP.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is substantially permeable to oxygen and is amicroporous membrane prepared from polysulfone. In certain aspects, thepolysulfone membrane is a hydrophobic microporous membrane that issubstantially permeable to oxygen.

In aspects according to the present disclosure, the microporouspolysulfone membrane comprises pores having a range of between 0.01 μmand 2.0 μm. In other aspects, the microporous polysulfone membrane 113comprises pores having a range of between 0.01 μm and 1.0 μm. In someaspects, a microporous polysulfone membrane 113 has a pore size ofbetween 0.03 μm and 1.0 μm in diameter. In other aspects, a microporouspolysulfone membrane 113 has a pore size of between 0.03 μm and 0.45 μmin diameter.

In aspects according to the present disclosure, the void fraction of apolysulfone membrane 113 used to prepare a collapsible blood container102 is between 20 and 80%. In another aspect, the void fraction of apolysulfone membrane 113 used to prepare a collapsible blood container102 is between 35 and 50%.

In certain aspects, the permeability polysulfone membranes havingmicropores greater than about 0.2 μm may allow fluid to permeate throughthe membrane, compromising both the fluid containment and the oxygen andcarbon dioxide permeability. To overcome this permeability at high poresizes, so called “super-hydrophobic” membranes can be employed whereinthe contact angle is greater than 150°. As used herein and known in theart, the contact angle quantifies the wettability of a solid surface andis theoretically described by Young's equation. In certain aspectsaccording the present disclosure, the use of non-hydrophobic polysulfonematerials is not recommended as the surface tension of the material islower and allows for fluid to seep through the pores even at the rangesstated above.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a polysulfone permeable membrane113 having a pore size of between 0.3 μm and 0.8 μm in diameter. Inother aspects, micropores of porous polysulfone membranes may be from0.22 μm to 0.8 μm in diameter. In an aspect, the micropores of porouspolysulfone membranes are from 0.2 μm to 1.0 μm. In another aspect, themicropores of porous polysulfone membranes may be greater than 0.1 μmand less than 1.0 μm. In a further aspect, the micropore of the porouspolysulfone membrane ranges from about 0.05 μm to about 1.0 μm. In someaspects, the micropores of porous polysulfone membranes may be greaterthan 0.3 μm or 0.4 μm. In other aspects, the micropores of porouspolysulfone membranes may be greater than 0.5 μm or 0.6 μm.

In aspects according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga polysulfone membrane 113 having a micropore size of less than 1.0 μm.In another aspect according to the present disclosure, an oxygendepletion device 10 comprises an inner collapsible blood container 102comprising a polysulfone membrane 113 having a micropore size of lessthan 0.8 μm. In certain aspects according to the present disclosure, anoxygen depletion device 10 comprises an inner collapsible bloodcontainer 102 comprising a polysulfone membrane 113 having a microporesize of less than 0.65 μm. In another aspect according to the presentdisclosure, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a polysulfone membrane 113 having amicropore size of less than 0.45 μm.

In an aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga polysulfone membrane 113 having a micropore size of 0.1 μm. In anotheraspect, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a polysulfone membrane 113 having amicropore size of 0.22 μm. In another aspect, an oxygen depletion device10 comprises an inner collapsible blood container 102 comprising apolysulfone membrane 113 having a micropore size of 0.20 μm. In afurther aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga polysulfone membrane 113 having a micropore size of 0.45 μm. In yet afurther aspect, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a polysulfone membrane 113having a micropore size of 0.65 μm. In another aspect according to thepresent disclosure, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a polysulfone membrane 113having a micropore size of 0.8 μm. In another aspect according to thepresent disclosure, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a polysulfone membrane 113having a micropore size of 0.03 μm. In another aspect according to thepresent disclosure, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a polysulfone membrane 113having a micropore size of 0.05 μm. In another aspect according to thepresent disclosure, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a polysulfone membrane 113having a micropore size of 1.2 μm.

In aspects according to the present disclosure, the polysulfone membranemay be less than 250 μm thick. In certain aspects, the membrane isgreater than 10 μm thick. In some aspects, the polysulfone membrane maybe between 10 and 250 μm thick. In other aspects, the polysulfonemembrane may be between 10 and 125 μm thick or 25 and 150 μm thick. Inan aspect, the polysulfone membrane may be between 50 and 125 μm thick,75 and 125 μm thick, 50 and 150 μm thick, 75 and 150 μm thick, 100 and125 μm thick, 150 and 250 μm thick, or between 25 and 150 μm thick. Inan aspect, the membrane 113 of inner collapsible blood container 102 isabout 20 μm thick. In another aspect, the membrane 113 of innercollapsible blood container 102 is about 30 μm thick. In yet anotheraspect, the membrane 113 of inner collapsible blood container 102 isabout 50 μm thick. In a further aspect, the membrane 113 of innercollapsible blood container 102 is about 76 μm thick. In an aspect, themembrane 113 of inner collapsible blood container 102 is about 120 μmthick.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a polysulfone permeable membrane113 that is between 100 and 125 μm thick. In certain aspects accordingto the present disclosure, the collapsible blood container 102 isprepared from a polysulfone permeable membrane 113 having a pore size ofbetween 0.1 μm and 0.8 μm in diameter and that is between 100 and 125 μmthick. In certain aspects according to the present disclosure, thecollapsible blood container 102 is prepared from a polysulfone permeablemembrane 113 having a pore size of between 0.1 μm and 0.8 μm in diameterand that is between 50 and 150 μm thick.

Examples of suitable polysulfone membranes for the preparation of innercollapsible blood containers that are permeable to oxygen according tothe present disclosure include SS003AH 10-250 μm thick/0.03 μm pore;SS005AH 10-250 μm thick/0.05 μm pore; SS010AH 10-250 μm thick/0.1 μmpore; SS020AH 10-250 μm thick/0.2 μm pore; SS045AH 10-250 μm thick/0.45μm pore; SS065AH 10-250 μm thick/0.65 μm pore; SS080AH 10-250 μmthick/0.8 μm pore; or SS120AH 10-250 μm thick/1.2 μm pore.

Suitable polysulfone membranes include commercially available membranes.Non-limiting examples of polysulfone membranes are available fromPacific Membranes. In an aspect, the polysulfone membrane may beSS120AH, SS080AH, SS065AH, SS045AH, SS020AH, SS010AH, SS005AH, orSS003AH.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is substantially permeable to oxygen and is amicroporous membrane prepared from polyolefin. In certain aspects, thepolyolefin membrane is a hydrophobic microporous membrane that issubstantially permeable to oxygen.

In aspects according to the present disclosure, the microporouspolyolefin membrane comprises pores having a range of between 0.01 μmand 2.0 μm. In other aspects, the microporous polyolefin membrane 113comprises pores having a range of between 0.01 μm and 1.0 μm. In someaspects, a microporous polyolefin membrane 113 has a pore size ofbetween 0.03 μm and 1.0 μm in diameter. In other aspects, a microporouspolyolefin membrane 113 has a pore size of between 0.03 μm and 0.45 μmin diameter.

In aspects according to the present disclosure, the void fraction of apolyolefin membrane 113 used to prepare a collapsible blood container102 is between 20 and 80%. In another aspect, the void fraction of apolyolefin membrane 113 used to prepare a collapsible blood container102 is between 35 and 50%.

In certain aspects, the permeability polyolefin membranes havingmicropores greater than about 1.0 μm may allow fluid to permeate throughthe membrane, compromising both the fluid containment and the oxygen andcarbon dioxide permeability. To overcome this permeability at high poresizes, so called “super-hydrophobic” membranes can be employed whereinthe contact angle is greater than 150°. As used herein and known in theart, the contact angle quantifies the wettability of a solid surface andis theoretically described by Young's equation. In certain aspectsaccording the present disclosure, the use of non-hydrophobic polyolefinmaterials is not recommended as the surface tension of the material islower and allows for fluid to seep through the pores even at the rangesstated above.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a polyolefin permeable membrane 113having a pore size of between 0.1 μm and 0.8 μm in diameter. In otheraspects, micropores of porous polyolefin membranes may be from 0.22 μmto 0.8 μm in diameter. In an aspect, the micropores of porous polyolefinmembranes are from 0.2 μm to 1.0 μm. In another aspect, the microporesof porous polyolefin membranes may be greater than 0.1 and less than 1.0μm. In a further aspect, the micropore of the porous polyolefin membraneranges from about 0.05 μm to about 1.0 μm. In some aspects, themicropores of porous polyolefin membranes may be greater than 0.3 or 0.4μm. In other aspects, the micropores of porous polyolefin membranes maybe greater than 0.5 or 0.6 μm.

In aspects according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga polyolefin membrane 113 having a micropore size of less than 1.0 μm.In another aspect according to the present disclosure, an oxygendepletion device 10 comprises an inner collapsible blood container 102comprising a polyolefin membrane 113 having a micropore size of lessthan 0.8 μm. In certain aspects according to the present disclosure, anoxygen depletion device 10 comprises an inner collapsible bloodcontainer 102 comprising a polyolefin membrane 113 having a microporesize of less than 0.65 μm. In another aspect according to the presentdisclosure, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a polyolefin membrane 113 having amicropore size of less than 0.45 μm.

In an aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga polyolefin membrane 113 having a micropore size of 0.1 μm. In anotheraspect, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a polyolefin membrane 113 having amicropore size of 0.22 μm. In another aspect, an oxygen depletion device10 comprises an inner collapsible blood container 102 comprising apolyolefin membrane 113 having a micropore size of 0.20 μm. In a furtheraspect according to the present disclosure, an oxygen depletion device10 comprises an inner collapsible blood container 102 comprising apolyolefin membrane 113 having a micropore size of 0.45 μm. In yet afurther aspect, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a polyolefin membrane 113having a micropore size of 0.65 μm. In another aspect according to thepresent disclosure, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a polyolefin membrane 113having a micropore size of 0.8 μm.

In aspects according to the present disclosure, the polyolefin membranemay be less than 250 μm thick. In certain aspects, the membrane isgreater than 10 μm thick. In some aspects, the polyolefin membrane maybe between 10 and 250 μm thick. In other aspects, the polyolefinmembrane may be between 10 and 125 μm thick or between 25 and 150 μmthick. In an aspect, the polyolefin membrane may be between 50 and 125μm thick, 75 and 125 μm thick, 50 and 150 μm thick, 75 and 150 μm thick,100 and 125 μm thick, 150 and 250 μm thick, or between 25 and 150 μmthick. In an aspect, the membrane 113 of inner collapsible bloodcontainer 102 is about 20 μm thick. In another aspect, the membrane 113of inner collapsible blood container 102 is about 30 μm thick. In yetanother aspect, the membrane 113 of inner collapsible blood container102 is about 50 μm thick. In a further aspect, the membrane 113 of innercollapsible blood container 102 is about 76 μm thick. In an aspect, themembrane 113 of inner collapsible blood container 102 is about 120 μmthick.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a polyolefin permeable membrane 113that is between 100 and 125 μm thick. In certain aspects according tothe present disclosure, the collapsible blood container 102 is preparedfrom a polyolefin permeable membrane 113 having a pore size of between0.1 μm and 0.8 μm in diameter and that is between 100 μm and 125 μmthick. In certain aspects according to the present disclosure, thecollapsible blood container 102 is prepared from a polyolefin permeablemembrane 113 having a pore size of between 0.1 μm and 0.8 μm in diameterand that is between 50 μm and 150 μm thick.

Examples of suitable polyolefin membranes for the preparation of innercollapsible blood containers that are permeable to oxygen according tothe present disclosure include those described in U.S. Pat. No.4,440,815 issued to Zomorodi et al.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is substantially permeable to oxygen and is amicroporous membrane prepared from polytetrafluoroethylene (PTFE). Incertain aspects, the PTFE membrane is a hydrophobic microporous membranethat is substantially permeable to oxygen.

In aspects according to the present disclosure, the microporous PTFEmembrane comprises pores having a range of between 0.01 μm and 2.0 μm.In other aspects, the microporous PTFE membrane 113 comprises poreshaving a range of between 0.01 μm and 1.0 μm. In some aspects, amicroporous PTFE membrane 113 has a pore size of between 0.03 μm and 1.0μm in diameter. In other aspects, a microporous PTFE membrane 113 has apore size of between 0.03 μm and 0.45 μm in diameter.

In aspects according to the present disclosure, the void fraction of aPTFE membrane 113 used to prepare a collapsible blood container 102 isbetween 20 and 80%. In another aspect, the void fraction of a PTFEmembrane 113 used to prepare a collapsible blood container 102 isbetween 35 and 50%.

In certain aspects, the permeability PTFE membranes having microporesgreater than about 1.0 μm may allow fluid to permeate through themembrane, compromising both the fluid containment and the oxygen andcarbon dioxide permeability. To overcome this permeability at high poresizes, so called “super-hydrophobic” membranes can be employed whereinthe contact angle is greater than 150°. As used herein and known in theart, the contact angle quantifies the wettability of a solid surface andis theoretically described by Young's equation. In certain aspectsaccording the present disclosure, the use of non-hydrophobic PTFEmaterials is not recommended as the surface tension of the material islower and allows for fluid to seep through the pores even at the rangesstated above.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a PTFE permeable membrane 113having a pore size of between 0.1 μm and 0.8 μm in diameter. In otheraspects, micropores of porous PTFE membranes may be from 0.22 μm to 0.8μm in diameter. In an aspect, the micropores of porous PTFE membranesare from 0.2 μm to 1.0 μm. In another aspect, the micropores of porousPTFE membranes may be greater than 0.1 and less than 1.0 μm. In afurther aspect, the micropore of the porous PTFE membrane ranges fromabout 0.05 μm to about 1.0 μm. In some aspects that micropores of porousPTFE membranes may be greater than 0.3 or 0.4 μm. In other aspects, themicropores of porous PTFE membranes may be greater than 0.5 or 0.6 μm.

In aspects according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PTFE membrane 113 having a micropore size of less than 1.0 μm. Inanother aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PTFE membrane 113 having a micropore size of less than 0.8 μm. Incertain aspects according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PTFE membrane 113 having a micropore size of less than 0.65 μm. Inanother aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PTFE membrane 113 having a micropore size of less than 0.45 μm.

In an aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga PTFE membrane 113 having a micropore size of 0.1 μm. In anotheraspect, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a PTFE membrane 113 having a microporesize of 0.22 μm. In another aspect, an oxygen depletion device 10comprises an inner collapsible blood container 102 comprising a PTFEmembrane 113 having a micropore size of 0.20 μm. In a further aspectaccording to the present disclosure, an oxygen depletion device 10comprises an inner collapsible blood container 102 comprising a PTFEmembrane 113 having a micropore size of 0.45 μm. In yet a furtheraspect, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a PTFE membrane 113 having a microporesize of 0.65 μm. In another aspect according to the present disclosure,an oxygen depletion device 10 comprises an inner collapsible bloodcontainer 102 comprising a PTFE membrane 113 having a micropore size of0.8 μm.

In aspects according to the present disclosure, the PTFE membrane 113may be less than 250 μm thick. In certain aspects, the membrane isgreater than 10 μm thick. In some aspects the PTFE membrane 113 may bebetween 10 and 250 μm thick. In other aspects, the PTFE membrane 113 maybe between 10 and 125 or 25 and 150 μm thick. In an aspect, the PTFEmembrane 113 may be between 50 and 125 μm thick, 75 and 125 μm thick, 50and 150 μm thick, 75 and 150 μm thick, 100 and 125 μm thick, 150 and 250μm thick or between 25 and 150 μm thick. In another aspect, the membrane113 of inner collapsible blood container 102 is about 30 μm. In yetanother aspect, the membrane 113 of inner collapsible blood container102 is about 50 μm. In a further aspect, the membrane 113 of innercollapsible blood container 102 is about 76 μm. In an aspect, themembrane 113 of inner collapsible blood container 102 is about 120 μmthick, 100 and 125 μm thick, 150 and 250 μm thick or between 25 and 150μm thick.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a PTFE permeable membrane 113 thatis between 100 and 125 μm thick. In certain aspects according to thepresent disclosure, the collapsible blood container 102 is prepared froma PTFE permeable membrane 113 having a pore size of between 0.1 μm and0.8 μm in diameter and that is between 100 and 125 μm thick. In certainaspects according to the present disclosure, the collapsible bloodcontainer 102 is prepared from a PTFE permeable membrane 113 having apore size of between 0.1 μm and 0.8 μm in diameter and that is between50 and 150 μm thick.

Examples of suitable PTFE membranes for the preparation of innercollapsible blood containers that are permeable to oxygen according tothe present disclosure include the Poreflon® FP, WP, and HP series PTFEmembranes from Sumitomo Electric Interconnect Products, San Marcos,Calif., and Tetratex® 2 from Donaldson Membranes, Ivyland, Pa.

Suitable PTFE membranes include commercially available membranes.Non-limiting examples of PTFE membranes are available from SumitomoElectric Interconnect Products, San Marcos, Calif., and DonaldsonMembranes, Ivyland, Pa. In an aspect, the PTFE membrane may be FP-010from Sumitomo Electric Interconnect Products, San Marcos, Calif.

In certain aspects, suitable membranes that are substantially permeableto oxygen may be multilayered membranes. In certain aspects, themultilayered membranes are hydrophobic microporous membranes that aresubstantially permeable to oxygen. Suitable multilayered membranesinclude multilayered membranes having two or more materials selectedfrom the group consisting of PVDF rendered hydrophobic, nylon, celluloseesters, polysulfone, polyethersulfone, polypropylene renderedhydrophobic, and polyacrylonitrile.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is substantially permeable to oxygen and is amicroporous membrane prepared from an extruded, woven, non-woven singlelayer or multilayered membrane. In certain aspects, the multilayeredmembrane is a hydrophobic microporous membrane that is substantiallypermeable to oxygen.

In aspects according to the present disclosure, the microporousmultilayered membrane comprises pores having a range of between 0.01micrometer (μm) and 2.0 μm. In other aspects, the microporousmultilayered membrane 113 comprises pores having a range of between 0.01μm and 1.0 μm. In some aspects, a microporous multilayered membrane 113has a pore size of between 0.03 μm and 1.0 μm in diameter. In otheraspects, a microporous multilayered membrane 113 has a pore size ofbetween 0.03 μm and 0.45 μm in diameter.

In aspects according to the present disclosure, the void fraction of amultilayered membrane 113 used to prepare a collapsible blood container102 is between 20 and 80%. In another aspect, the void fraction of amultilayered membrane 113 used to prepare a collapsible blood container102 is between 35 and 50%.

In certain aspects, the permeability of multilayered membranes havingmicropores greater than about 1.0 μm may allow fluid to permeate throughthe membrane, compromising both the fluid containment and the oxygen andcarbon dioxide permeability. To overcome this permeability at high poresizes, so called “super-hydrophobic” membranes can be employed whereinthe contact angle is greater than 150°. As used herein and known in theart, the contact angle quantifies the wettability of a solid surface andis theoretically described by Young's equation. In certain aspectsaccording the present disclosure, the use of non-hydrophobicmultilayered materials is not recommended as the surface tension of thematerial is lower and allows for fluid to seep through the pores even atthe ranges stated above.

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a multilayered permeable membrane113 having a pore size of between 0.1 μm and 0.8 μm in diameter. Inother aspects, micropores of porous multilayered membranes may be from0.22 μm to 0.8 μm in diameter. In an aspect, the micropores of porousmultilayered membranes are from 0.2 and to 1.0 μm. In another aspect,the micropores of porous multilayered membranes may be greater than 0.1and less than 1.0 μm. In a further aspect, the micropore of the porousmultilayered membrane ranges from about 0.05 μm to about 1.0 μm. In someaspects, the micropores of porous multilayered membranes may be greaterthan 0.3 or 0.4 μm. In other aspects, the micropores of porousmultilayered membranes may be greater than 0.5 or 0.6 μm.

In aspects according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga multilayered membrane 113 having a micropore size of less than 1.0 μm.In another aspect according to the present disclosure, an oxygendepletion device 10 comprises an inner collapsible blood container 102comprising a multilayered membrane 113 having a micropore size of lessthan 0.8 μm. In certain aspects according to the present disclosure, anoxygen depletion device 10 comprises an inner collapsible bloodcontainer 102 comprising a multilayered membrane 113 having a microporesize of less than 0.65 μm. In another aspect according to the presentdisclosure, an oxygen depletion device 10 comprises an inner collapsibleblood container 102 comprising a multilayered membrane 113 having amicropore size of less than 0.45 μm.

In an aspect according to the present disclosure, an oxygen depletiondevice 10 comprises an inner collapsible blood container 102 comprisinga multilayered membrane 113 having a micropore size of 0.1 μm. Inanother aspect, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a multilayered membrane 113having a micropore size of 0.22 μm. In another aspect, an oxygendepletion device 10 comprises an inner collapsible blood container 102comprising a multilayered membrane 113 having a micropore size of 0.20μm. In a further aspect according to the present disclosure, an oxygendepletion device 10 comprises an inner collapsible blood container 102comprising a multilayered membrane 113 having a micropore size of 0.45μm. In yet a further aspect, an oxygen depletion device 10 comprises aninner collapsible blood container 102 comprising a multilayered membrane113 having a micropore size of 0.65 μm. In another aspect according tothe present disclosure, an oxygen depletion device 10 comprises an innercollapsible blood container 102 comprising a multilayered membrane 113having a micropore size of 0.8 μm.

In aspects according to the present disclosure, the multilayeredmembrane 113 may be less than 250 μm thick. In certain aspects, themembrane is greater than 10 μm thick. In some aspects the multilayeredmembrane 113 may be between 10 and 250 μm thick. In other aspects, themultilayered membrane may be between 10 and 125 μm thick or 25 and 150μm thick. In an aspect, the multilayered membrane 113 may be between 50and 125 μm thick, 75 and 125 μm thick, 50 and 150 μm thick, 75 and 150μm thick, 100 and 125 μm thick, 150 and 250 μm thick or between 25 and150 μm thick, 100 and 125 μm thick, 150 and 250 μm thick or between 25and 150 μm thick. In another aspect, the membrane 113 of innercollapsible blood container 102 is about 30 μm. In yet another aspect,the membrane 113 of inner collapsible blood container 102 is about 50μm. In a further aspect, the membrane 113 of inner collapsible bloodcontainer 102 is about 76 μm. In an aspect, the membrane 113 of innercollapsible blood container 102 is about 120 μm thick

In certain aspects according to the present disclosure, the collapsibleblood container 102 is prepared from a multilayered permeable membrane113 that is between 100 and 125 μm thick. In certain aspects accordingto the present disclosure, the collapsible blood container 102 isprepared from a multilayered permeable membrane 113 having a pore sizeof between 0.1 μm and 0.8 μm in diameter and that is between 100 μm and125 μm thick. In certain aspects according to the present disclosure,the collapsible blood container 102 is prepared from a multilayeredpermeable membrane 113 having a pore size of between 0.1 μm and 0.8 μmin diameter and that is between 50 μm and 150 μm thick.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is substantially permeable to oxygen and is amembrane prepared from polyvinyl chloride (PVC). In aspects accordingthe present disclosure, the collapsible blood container 102 can beprepared from a PVC membrane having a thickness of between 5 μm and 250μm, and more preferably between about 10 μm and about 100 μm.

The use of PVC in the manufacture of collapsible blood containers iswell known in the art. The use of various plasticizers in various PVCformulations is also well known in the art, and includes the use ofdiethylhexyl phthalate (DEHP) for long term storage of red blood cells.Typical manufacture of collapsible blood containers from PVC-DEHPutilizes radiofrequency (RF) welding of a pair of films to convenientlyfabricate a bag structure, with such individual films having a thicknessof about 350 μm to about 400 μm. An exemplary PVC-DEHP film is theRenolit ES-3000 film (American Renolit Corp., City of Commerce, Calif.).

Due to the relatively low oxygen permeability of such films and the needfor higher oxygen permeability for platelet storage, other plasticizersfor PVC have found utility in the fabrication of collapsible bloodcontainers and include the use of citrate, among others (see, forexample, “The Role of Poly(Vinyl Chloride) in Healthcare” by Colin R.Blass, copyright 2001 Rapra Technology, Ltd., ISBN:1-85957-258-8). Asuitable example of a PVC-citrate film is the Renolit ES-4000 film(American Renolit Corp., City of Commerce, Calif.).

The present disclosure provides for suitable PVC materials for use in acollapsible blood container 102 that is substantially permeable tooxygen. The use of a PVC-citrate film such as Renolit ES-4000 having athickness of from about 5 μm to about 250 μm, and more preferably fromabout 10 μm to about 100 μm is suitable for providing a collapsibleblood container having the desired characteristics of high oxygenpermeability, RF welding and joining, and high tensile strength.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 that is substantially permeable to oxygen and is amembrane prepared from silicone. In aspects according the presentdisclosure, the collapsible blood container 102 can be prepared from asilicone membrane having a thickness of between 15 μm and 100 μm. Inaspects according the present disclosure, the collapsible bloodcontainer 102 can be prepared from a silicone membrane having athickness of between 5 μm and 500 μm. In other aspects, the collapsibleblood container 102 can have a thickness of between 5 μm and 200 μm. Inother aspects, the collapsible blood container 102 can have a thicknessof between 20 μm and 120 μm. In another aspect the collapsible bloodcontainer 102 is between 30 μm and 120 μm thick. In yet another aspect,the collapsible blood container 102 is between 50 μm and 120 μm thick.In a further aspect, the thickness of the collapsible blood container102 can be between 76 μm and 120 μm. In another aspect the collapsibleblood container 102 is between 20 μm and 50 μm thick. The presentdisclosure provides for, and includes, a collapsible blood container 102that is 20 μm in thickness. In another aspect, the collapsible bloodcontainer 102 is 15 μm thick. In another aspect, the collapsible bloodcontainer 102 is 30 μm thick. In yet another aspect, the collapsibleblood container 102 is 50 μm thick. In an additional aspect, thecollapsible blood container 102 is 120 μm thick.

In aspects according the present disclosure, the collapsible bloodcontainer 102 can be prepared from a silicone membrane having athickness of between 20 μm and 400 μm. In other aspects, the collapsibleblood container 102 can have a thickness of between 20 μm and 200 μm. Inother aspects, the collapsible blood container 102 can have a thicknessof between 40 μm and 300 μm. In another aspect, the collapsible bloodcontainer 102 is between 40 μm and 400 μm thick. In yet another aspect,the collapsible blood container 102 is between 300 μm and 450 μm thick.In a further aspect, the thickness of the collapsible blood container102 can be between 350 μm and 450 μm. The present disclosure providesfor, and includes, a collapsible blood container 102 that is about 450μm in thickness. In another aspect, the collapsible blood container 102is 425 μm thick. In yet another aspect, the collapsible blood container102 is 400 μm thick. In an additional aspect, the collapsible bloodcontainer 102 is 350 μm thick.

Suitable silicone membranes include commercially available membranes.Non-limiting examples of silicone membranes are available from WackerSilicones, such as the Silpuran® brand of medical grade silicone sheetmembranes (Wacker Silicones, Adrian, Mich.) and Polymer Sciences PS-1033P-Derm® silicone elastomer membrane (Polymer Sciences, Inc., Monticello,Ind.). In an aspect, the silicone membrane may be Polymer SciencesPS-1033 or Wacker Silpuran® 6000 silicone. Silicone membranes can beprepared from various liquid silicone rubber (LSR) materials, which areavailable from a number of silicone suppliers, such as Wacker Silicones(Adrian, Mich.), Shin-Etsu Silicones of America (Akron, Ohio), NuSilTechnology (Carpenteria, Calif.), and Blue Star Silicones (EastBrunswick, N.J.), to name a few.

In an aspect according to the present disclosure, a collapsible bloodcontainer 102 can be manufactured from silicone by various moldingmethods such as compression molding, injection molding, and insertmolding, and also adhesive bonding of silicone sheets using siliconeadhesives. In one aspect according to the present disclosure, a pair ofsilicone sheets are bonded together around the periphery with a sectionof silicone inlet tubing in place in the seam using silicone adhesive.In another aspect according to the present disclosure, a silicone liquidrubber is injection molded over a form to create a three-sided shape,which is then further bonded to closure on the remaining fourth sidearound a silicone inlet tube using a silicone adhesive. In anotheraspect according to the present disclosure, a silicone liquid rubber isinjection molded over a form to create a three-sided shape, which isthen insert molded onto a closure shape on the remaining fourth sidethat incorporates an inlet tubing into the closure shape.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 having resistance to tearing. As used herein, “tearresistance” or “tear strength” is measured in kN/m. In aspects accordingthe present disclosure, the collapsible blood container 102 should beprepared from oxygen permeable materials that are also resistant totearing. Measures of tear resistance are known in the art, for example,ASTM D-412, which can also be used to measure tensile strength, modulus,and elongations. In certain aspects, collapsible blood container 102should be prepared from oxygen permeable materials that are resistant tothe formation of a tear (e.g., tear initiation). Methods of measuringtear initiation and tear propagation are known in the art, for exampleASTM D-624. Other methods include measuring the tensile strength and theelongation at break according to DIN 53 504-S1.

In an aspect according to the present disclosure, a collapsible bloodcontainer 102 should be prepared from oxygen permeable materials havinga tensile strength of at least 2.4 N/mm².

The present disclosure provides for, and includes, sorbents capable ofbinding to and removing oxygen from an environment. Unless providedotherwise, the term “sorbent” refers to oxygen sorbents and scavengers.As used herein, “oxygen scavenger” or “oxygen sorbent” is a materialthat binds irreversibly to or combines with O₂ under the conditions ofuse. The term “oxygen sorbent” may be used interchangeably herein with“oxygen scavenger.” In certain aspects according the present disclosure,a material may bind to or combines with oxygen irreversibly. In otheraspects, oxygen may bind to a sorbent material and have a very slow rateof release, k_(off). In an aspect, the oxygen may chemically react withsome component of the material and be converted into another compound.Any material where the off-rate of bound oxygen is much less than theresidence time of the blood can serve as an oxygen scavenger.

As used herein, the amount of sorbent is provided as having a certainbinding capacity of oxygen as measured by volume (e.g., cubiccentimeters (cc) or milliliters (ml)) at standard temperature andpressure (e.g., 0° C. (273.15 Kelvin) and 1.01×10⁵ pa (100 kPa, 1 bar,0.986 atm, 760 mmHg) of pressure). In other aspects, oxygen sorbents andscavengers are further capable of binding to and removing carbon dioxidefrom an environment. In certain aspects, sorbent 103 may be a mixture ofnon-toxic inorganic and/or organic salts and ferrous iron or othermaterials with high reactivity toward oxygen, carbon dioxide, or oxygenand carbon dioxide. In certain aspects, an oxygen sorbent or scavengeris combined with a carbon dioxide sorbent. In other aspects, thepresence or absence of carbon dioxide binding capabilities of an oxygensorbent is not necessary.

Suitable oxygen sorbents or scavengers are known in the art. Suitableoxygen sorbents according to the present disclosure have minimum oxygenadsorption rates of 0.44 ml/min. Sorbents having suitable adsorptionprofiles bind at least 45 ml O₂ within 60 minutes, 70 ml O₂ within 120minutes, and 80 ml O₂ within 180 minutes. Suitable sorbents may haveboth higher capacity and binding rates.

Non-limiting examples of oxygen scavengers or sorbents include ironpowders and organic compounds. Examples of O₂ sorbents include chelatesof cobalt, iron, and Schiff bases. Additional non-limiting examples forO₂ sorbents may be found in U.S. Pat. No. 7,347,887 issued to Bulow etal., U.S. Pat. No. 5,208,335, issued to Ramprasad et al., and U.S. Pat.No. 4,654,053 issued to Sievers et al.; each of which is herebyincorporated by reference in their entireties. Oxygen sorbent materialsmay be formed into or incorporated in fibers, microfibers, microspheres,microparticles, and foams.

In certain aspects, suitable sorbents include those obtainable fromMultisorb Technologies (Buffalo, N.Y.), Sorbent Systems/ImpakCorporation (Los Angeles, Calif.) or Mitsubishi Gas Chemical America(MGC) (New York, N.Y.). Exemplary oxygen sorbents include MultisorbTechnologies StabilOx® packets, Sorbent Systems P/N SF100PK100 100 ccoxygen absorber, and Mitsubishi Gas Chemical America Ageless® SS-200oxygen absorber. MGC also provides sorbents suitable for the methods anddevices of the present disclosure. Such suitable oxygen sorbents includethe MGC Ageless® and SS-200 oxygen absorber.

In aspects according to the present disclosure, a sorbent may be anoxidizable organic polymer having a polymeric backbone and a pluralityof pendant groups. Examples of sorbents with a polymeric backboneinclude a saturated hydrocarbon (<0.01% carbon-carbon double bonds). Insome aspects, the backbone can contain monomers of ethylene or styrene.In an aspect, a polymeric backbone may be ethylenic. In another aspect,an oxidizable organic compound may be ethylene/vinyl cyclohexenecopolymer (EVCH). Additional examples of substituted moieties andcatalysts are provided in U.S. Patent Publication No. 2003/0183801 byYang et al., hereby incorporated by reference in its entirety. Inadditional aspects, an oxidizable organic polymer can also comprisesubstituted hydrocarbon moieties. Examples of oxygen scavenging polymersinclude those described by Ching et al., International PatentPublication W099/48963, hereby incorporated by reference in itsentirety. Oxygen scavenging materials may include those provided in U.S.Pat. No. 7,754,798 issued to Ebner et al., U.S. Pat. No. 7,452,601issued to Ebner et al., or U.S. Pat. No. 6,387,461 issued to Ebner etal.; each of which are hereby incorporated by reference in theirentireties.

As used herein, sorbents of the present disclosure may be either free orcontained in a permeable enclosure, container, envelope, etc. In certainaspects, sorbent is provided in one or more sachets made of materialshaving high porosity and essentially no resistance to the transport ofgases. Examples of such materials include spun polyester films,perforated metallic foils, and combinations thereof.

The present disclosure further includes, and provides for, sorbentincorporated as one or more laminated layers of an outer articlesubstantially impermeable to oxygen. Polymeric sorbents such as thosedescribed above may be laminated to sheets used to prepare an outerreceptacle using methods known in the art, including soft contactlamination, thermal lamination, or solvent lamination.

The present disclosure further includes, and provides for, sorbentsformed inside the pores of porous micro-glass fibers or encapsulated inother inert materials. The encapsulation of transition-metal complexeswithin the pores of a porous material may be achieved by using aship-in-a-bottle synthesis in which the final molecule is preparedinside the pores by reacting smaller precursors. Examples of suchencapsulated sorbents are known in the art, for example, as described byKuraoka, et al., “Ship-in-a-bottle synthesis of a cobaltphthalocyanine/porous glass composite membrane for oxygen separation,”Journal of Membrane Science, 286(1-2):12-14 (2006), herein incorporatedby reference in its entirety. In some aspects, porous glass fibers maybe manufactured as provided in U.S. Pat. No. 4,748,121 issued to Beaveret al., herein incorporated by reference in its entirety. In anotheraspect, a sorbent can formed as a porous sheet product usingpapermaking/non-woven wet-laid equipment. Sheets with O₂ scavengingformulations may be as described in U.S. Pat. No. 4,769,175 issued toInoue, herein incorporated by reference in its entirety, which can beformed and then encapsulated with a silicone film.

As used herein, “carbon dioxide scavenger” is a material that binds toor combines with carbon dioxide under the conditions of use. The term“carbon dioxide sorbent” may be used interchangeably herein with “carbondioxide scavenger.” In certain aspects, carbon dioxide sorbents may benon-reactive, or minimally reactive with oxygen. In other embodiments,oxygen sorbents may exhibit a secondary functionality of carbon dioxidescavenging. Carbon dioxide scavengers include metal oxides and metalhydroxides. Metal oxides react with water to produce metal hydroxides.The metal hydroxide reacts with carbon dioxide to form water and a metalcarbonate. In certain aspects according the present disclosure, amaterial may bind to or combine with CO₂ irreversibly. In aspectsaccording to the present disclosure, a material may bind CO₂ with higheraffinity than hemoglobin. In other aspects, a sorbent material may bindCO₂ with high affinity such that the carbonic acid present in the bloodor RBC cytoplasm is released and absorbed by the sorbent. In otheraspects, CO₂ binds to a sorbent material and has a very slow rate ofrelease, k_(off). In an aspect, the carbon dioxide can chemically reactwith some component of the material and be converted into anothercompound.

Carbon dioxide scavengers are known in the art. In certain aspectsaccording to the present disclosure, a carbon dioxide scavenger may becalcium oxide. Reaction of calcium oxide with water produces calciumhydroxide that may react with carbon dioxide to form calcium carbonateand water. In certain aspects according the present disclosure, thewater for the production of calcium hydroxide is obtained via diffusionof blood derived water vapor through the inner oxygen permeablecontainer. In another aspect, the water may be provided by theenvironment through the outer receptacle that is substantiallyimpermeable to oxygen. In yet another aspect, the water may be includedwith the outer receptacle of the oxygen depletion device.

Non-limiting examples of CO₂ scavengers include oxygen scavengers andcarbon dioxide scavengers provided by Multisorb Technologies (Buffalo,N.Y.). Oxygen scavengers may exhibit a secondary functionality of carbondioxide scavenging.

In aspects according to the present disclosure, O₂ depletion media andCO₂ depletion media may be blended to a desired ratio to achieve desiredresults.

The present disclosure further includes and provides for sorbentscontained in sachets. As used herein, a “sachet” is any enclosure thatencloses and contains an oxygen sorbent, a carbon dioxide sorbent, or acombination of oxygen and carbon dioxide sorbent(s). Sachets accordingthe present disclosure are contained within overwrap material that isboth oxygen and carbon dioxide permeable. In certain embodiments, theoverwrap material may be a combination of two or more materials, atleast one of the materials being oxygen and carbon dioxide permeable.Suitable overwrap materials have a known biocompatible profile or meetISO 10993.

Sachets are sealed so that the sorbent contents are wholly containedwithin the overwrap material and do not allow the sorbent to leak orotherwise exit its overwrap package. Sachets may take any shape, thoughtypically take a rectangular or square shape. In an aspect, the sachetis about 50×60 mm. In an aspect, the oxygen sorbent 103 binds 30 ccoxygen per sachet at STP. In an aspect, the oxygen sorbent 103 binds 60cc oxygen per sachet at STP. In an aspect, the oxygen sorbent 103 binds120 cc oxygen per sachet at STP. In an aspect, the oxygen sorbent 103binds from 30 to 120 cc oxygen per sachet at STP. In an aspect, theoxygen sorbent 103 binds from 30 to 120 cc oxygen per sachet at STP. Inan aspect, the oxygen sorbent 103 binds from 50 to 200 cc oxygen persachet at STP. In certain aspects according to the present disclosure, asachet has a total oxygen adsorption capacity of 100 cc O₂ at STP. Incertain other aspects of the present disclosure, a sachet has a totaloxygen absorption capacity of at least 200 cc O₂ at STP.

In aspects according to the present disclosure, the oxygen sorbent 103may be provided in one or more sachets. In another aspect, the oxygensorbent 103 is provide in a single larger sachet. In other aspects, theoxygen sorbent 103 is provided in two sachets distributed within theheadspace between the inner collapsible container 102 and the outerreceptacle 101. In yet other aspects, the oxygen sorbent 103 is providedin four sachets distributed within the headspace between the innercollapsible container 102 and the outer receptacle 101. In aspectsaccording to the present disclosure, an oxygen depletion device 10 maycomprise 2 to 20 sorbent packages.

In aspects according to the present disclosure, oxygen depletion device10 includes from 1 to 50 grams of sorbent 103 contained in one or moresachets. In an aspect, an oxygen depletion device 10 includes from 1 to100 grams of sorbent 103 contained in one or more sachets. In an aspect,an oxygen depletion device 10 includes from 25 to 75 grams of sorbent103 contained in one or more sachets. In a further aspect, an oxygendepletion device 10 includes about 25 grams of sorbent 103. In yetanother aspect, oxygen depletion device 10 includes about 50 grams ofsorbent 103. In an aspect, an oxygen depletion device 10 includes about35 or 45 grams of sorbent 103 contained in one or more sachets. In anaspect, an oxygen depletion device 10 includes about 10 or 15 grams ofsorbent 103 contained in one or more sachets. The sachets can be square,rectangular, circular, or elliptical and have a perimeter of 40 to 150mm.

Sachets according to the present disclosure may further include a carbondioxide sorbent. In an aspect, an oxygen sorbent 103 also provides forcarbon dioxide adsorption. In an aspect, the oxygen sorbent 103 binds 30cc carbon dioxide at STP. In an aspect, the oxygen sorbent 103 binds atleast 170 cc oxygen and at least 30 cc carbon dioxide, where both gasesare measured at STP.

The present disclosure provides for, and includes, an outer receptacle101 that is substantially impermeable to oxygen. As discussed above, theintegrity of the oxygen barrier should be maintained when joining,welding, folding, or otherwise assembling an outer receptacle 101.Failures in assembly of the outer receptacle 101 compromises the shelflife of an oxygen depletion device 10 or renders it unable to performits intended purpose of depleting oxygen from blood. Importantly, bloodthat is inadequately depleted of oxygen does not realize the benefits ofdepletion during storage and may have significant negative consequenceswhen transfused into a patient. In addition to satisfying therequirements for blood collection and depletion, it is routine for bloodto be sampled through standardized ports 303 as well as for variousadditives to be introduced into the collected blood. More specifically,nearly all collected blood is provided with an anticoagulant at orduring collection.

To address the need to introduce materials into the collected blood, andto provide for the transfer of blood that has been depleted of oxygen toan appropriate anaerobic storage bag, an oxygen depletion device 10 mayfurther include one or more inlets/outlets 30. As provided herein,special care in the assembly of the outer receptacle 101 (and outerreceptacle 201) is necessary to ensure that when the oxygen impermeableouter receptacle 101 (and outer receptacle 201) is traversed, theinlet/outlet 30 does not become a source of unwanted oxygen ingress.

In aspects according to the present disclosure, the outer receptacle 101includes one or more inlets/outlets 30. In certain aspects, the one ormore inlet/outlets 30 further comprise a spike port 303.

It is notable that few materials provide complete impermeability andthat even the high impermeability of materials can be compromised whenjoining, welding, folding, or otherwise assembling an outer receptacle101. As will be discussed below, oxygen depletion device 10 may furtherincorporate optional spike ports 303 and inlets/outlets 30 and must alsobe designed to accommodate changes in volume of the inner collapsibleblood container 102. Accordingly, special care is taken to incorporatespecific design elements and manufacturing methods to ensure theintegrity of the impermeable barrier.

Spike ports 303 for use in blood collection kits and systems arecommonly known in the art and include products such as Vitalmed #20391(Vitalmed, Inc., Lakeville, Mass.) and Qosina 65842 (Qosina Corp.,Edgewood, N.Y.). These ports are typically molded from PVC and have aremovable cap that provides for a sterile barrier before use, and alsoprovides for some degree of oxygen impermeability to the contents. Insome aspects, a spike port 303 is covered by a sealed, frangible sectionof the outer receptacle film, thereby providing for a sterile barrierand also providing an additional degree of oxygen impermeability.Improved oxygen impermeability is desirable as it increases the shelflife of kits and systems having an oxygen depletion device 10.

As will be appreciated, conventional ports, inlets, and outlets arepotential sources of unwanted oxygen ingression that depend both on theselection of the material and the methods used to bond the port, inlet,or outlet to the outer receptacle 101. Methods of bonding materials arewell known in the art. As provided herein, inlet/outlet 30 comprises atube 301 joined to the outer receptacle 101 (or outer receptacle 201)using bond 302 which creates an oxygen impermeable seal to the outerreceptacle 101 (or outer receptacle 201). In an aspect, bond 302 isachieved by using constant heat sealing dies heated to and maintained atabout 210° F. In an aspect, films are placed between heated dies andclamped together for about to achieve a thermally welded seam. Incertain aspects, a heat seal is created in about 5 seconds. In certainaspects, the sealing dies have a grooved section machined out of them toaccommodate an intermediary component. In some aspects tube 301comprises an intermediary component that may be a length of multilayertubing as discussed below or a small block of machined polymer or moldeddevice. In certain aspects, a molded device is prepared from apolyolefin, such as polyethylene. In aspects according to the presentdisclosure, the groove is dimensioned about 10% smaller than thefeatures of the component, thereby providing for compression duringsealing.

In some aspects, an oxygen impermeable bond is comprised of a section ofmultilayer tube that is heat sealed into the seam of the outerreceptacle 101. In certain aspects, the multilayer tube is comprised ofan outer layer of polyethylene, and inner layer of PVC (polyvinylchloride), and an intermediary layer of EVA (ethyl-vinyl-alcohol)(Pexco, Inc. Athol, Mass.). In some aspects, additional sections of PVCtubing are solvent bonded into the multilayer tube using, for example,cyclohexanone.

In some aspects, an inlet/outlet 30 is comprised of a tube 301 preparedfrom a small diamond shaped block of polyethylene with a hole throughthe center, such that the diamond shaped block is heat sealed into theseam of the outer receptacle to provide an oxygen impermeable bond 302while the center through-hole provides for fluid connectivity with thecontents. In an aspect, a section of PVC tubing is bonded into thecenter hole of the diamond shaped block using an oxygen impermeableadhesive capable of bonding to polyethylene, such as Loctite 4310,Masterbond X17, or 3M Scothweld 4693, thereby providing for fluidconnectivity through the oxygen impermeable outer receptacle to thecontents therein. In other aspects, a multilayered tubing can be bondedto the center hole of the diamond shaped block using methods known inthe art. In other aspects, a multilayered tubing can be utilized inplace of standard PVC intravenous tubing to provide for enhanced oxygenbarrier properties.

The users of the collapsible container require convenient filling andremoval of the contents, and must be able to empty the contents within 2minutes per the ISO 3826 standard for blood containers. The outerreceptacle can reduce the filling time by constraining the collapsiblecontainer and preventing it from expanding. Thus, in some embodiments,the blood storage device is further comprised of an expansion feature toallow for unrestricted filling of the collapsible container. In someembodiments the expansion feature is comprised of a gusseted fold alongone or more edges of the outer receptacle. Typically, a fold of about ¼inch is adequate to provide for expansion of the inner container, andthe pleats of the fold are sealed into the seams at the ends. In someembodiments, the expansion feature is comprised of a third panel ofbarrier film sealed along the bottom of the outer receptacle, providingfor a three-dimensional bag.

During the development of the oxygen depletion device 10, it wasdiscovered that the size, shape, and number of chambers of an innercollapsible blood container 102 needed to be controlled in order toobtain suitable depletion kinetics. More particularly, even using highlypermeable materials, using standard blood bag configurations provedinadequate and had significantly slower reaction kinetics. Not to belimited by theory, it is hypothesized that deoxygenation is a multistepprocess including release of dissolved oxygen from hemoglobin, diffusionof the dissolved oxygen within the red blood cell cytoplasm, anddiffusion of the dissolved oxygen through the red blood cell membrane.Also not to be limited by theory, it is hypothesized that the highconcentration of hemoglobin, having very high affinity for oxygen,greatly decreases the diffusion rate of the dissolved oxygen within thecytoplasm. Similarly, the diffusion of dissolved oxygen once it passesthrough the plasma membrane to the plasma is further limited byabsorption and binding to other red cells. Again, not to be limited bytheory, it is hypothesized that an additional diffusion barrier for thedissolved oxygen occurs at the gas permeable membrane where it not onlyneeds to pass through the membrane, but also changes state from thedissolved phase to the gaseous phase. Subsequent diffusion andadsorption by the sorbent occurs in a gaseous state and is maximized byincorporating and maintaining a headspace within the outer receptacle101. Accordingly, it is believed that the diffusion of the gaseousoxygen is maximized by maintaining the concentration gradient within theheadspace from the surface of the inner collapsible blood container 102to the oxygen sorbent 103. Also not to be limited by theory, it isthought that by selecting sorbents that have high absorption kinetics,high binding capacity, and combinations of both, a suitable diffusiongradient for the gaseous oxygen is maintained to drive the rapidkinetics of oxygen depletion in oxygen depletion device 10.

The present disclosure provides for, and includes, an oxygen depletiondevice 10 for depleting oxygen from blood that comprises an innercollapsible blood container 102 having a surface to volume ratio ofbetween 4.75 centimeters²/milliliter (cm²/ml) and 6.9 cm²/ml enclosedwithin an outer receptacle 101. In certain aspects, an oxygen depletiondevice 10 for depleting oxygen from blood comprises an inner collapsibleblood container 102 having a surface to volume ratio of between 4.84cm²/ml and 6.9 cm²/ml enclosed within an outer receptacle 101 whenfilled with blood for oxygen depletion. In certain aspects, an oxygendepletion device 10 for depleting oxygen from blood comprises an innercollapsible blood container 102 having a surface to volume ratio ofbetween 5.0 cm²/ml and 6.9 cm²/ml enclosed within an outer receptacle101 when filled with blood for oxygen depletion. In some aspects, anoxygen depletion device 10 for depleting oxygen from blood comprises aninner collapsible blood container 102 having a surface to volume ratioof between 5.0 cm²/ml and 6.5 cm²/ml enclosed within an outer receptacle101 when filled with blood for oxygen depletion. In some aspects, anoxygen depletion device 10 for depleting oxygen from blood comprises aninner collapsible blood container 102 having a surface to volume ratioof between 5.5 cm²/ml and 6.5 cm²/ml enclosed within an outer receptacle101 when filled with blood for oxygen depletion.

As used herein, surface to volume and surface area to volume are usedinterchangeably throughout the present disclosure. A used herein,surface to volume ratios are defined with respect to a standard unit ofwhole blood, about 1 pint or 450-500 ml. As is evident to a person ofskill in the art, collection of less than a unit of blood results in aneven higher surface to volume ratio and the oxygen depletion device 10is suitable for collecting a fraction of a unit of blood withoutmodification. For the collection of more than a unit of blood, the sizeof the collapsible blood container 102 would need to be adjusted toprovide for the desirable rapid kinetics of blood depletion.Modifications of the sort necessary to adapt an oxygen depletion device10 for the collection of more than a unit of blood is within the levelof ordinary skill in the art.

The present disclosure further includes and provides for oxygendepletion device 10 for the collection and depletion of packed red bloodcells. A full unit of packed red blood cells in an additive solutioncomprises about 280±60 ml.

In an aspect according to the present disclosure, the surface to volumeratio of a collapsible blood container 102 is at least 4.84centimeters²/milliliter (cm²/ml) when filled with blood for oxygendepletion. Not to be limited by theory, it is believed that byincreasing the surface to volume ratio, the diffusion limitationsimposed by blood itself, particularly by the red blood cells andhemoglobin, can be overcome by decreasing the diffusion distance of thedissolved oxygen within the inner collapsible blood container 102. In anaspect, the surface to volume ratio of a blood container 102 is at least5.0 cm²/ml when filled with blood for oxygen depletion. In anotheraspect, the surface to volume ratio of a collapsible blood container 102is at least 5.5 cm²/ml when filled with blood for oxygen depletion. In afurther aspect, the surface to volume ratio of a collapsible bloodcontainer 102 is at least 6.0 cm²/ml when filled with blood for oxygendepletion. In some aspects, the surface to volume ratio of a collapsibleblood container 102 is at least 6.5 cm²/ml when filled with blood foroxygen depletion.

The present disclosure also includes and provides for increasing thekinetics of deoxygenation of blood by modifying the dimensions of theinner collapsible blood container 102. Not to be limited by theory, theaverage diffusion distance of a red blood cell in blood minimized as theheight is decreased leading to increased deoxygenation kinetics. Incertain aspects according the present disclosure, the collapsible bloodcontainer 102 is 25.4 cm by 30.5 cm by 0.02 cm before filling withblood, and about 1.5 cm in height after filling with blood. In otheraspects according the present disclosure, the collapsible bloodcontainer 102 is 17.5 cm by 28.0 cm (7×11 inches) by 0.04 cm beforefilling with blood, and about 2.0 cm in height after filling with blood.In other aspects according the present disclosure, the collapsible bloodcontainer 102 is 25.0 cm by 60.0 (10×23 inches) cm by 0.04 cm beforefilling with blood, and about 0.3 cm in height after filling with blood.

In certain aspects, the height of a collapsible blood container 102 isno greater than 0.005 cm when empty. In an aspect the height of acollapsible blood container 102 is no greater than 0.1 cm. In certainaspects, the height of a collapsible blood container 102 is between0.002 and 0.1 cm. When filled with blood, the height of a collapsibleblood container 102 is no greater than 0.3 cm. In an aspect the heightof a collapsible blood container 102 when filled with blood is nogreater than 1.5 cm. In certain aspects, the height of a collapsibleblood container 102 when filled with blood is between 0.2 cm and 2.5 cm.

The present disclosure also includes and provides for an oxygendepletion device 10 having dimensions suitable for incorporation ofexisting blood collection protocols using existing equipment. Design ofan oxygen depletion device 10 with recognition to existing technologiesreduces capital costs in centralized processing centers and furtherprovides for increased consistency and reliability. As used herein, thedimensions of an oxygen depletion device 10 is primarily limited to thelength and width of the outer receptacle 101 where the height of the bagis determined by the requirements of the collapsible blood container 102to contain about a pint or 450 to 500 ml of whole blood, which isequivalent to a “unit of blood”. In another aspect, the dimensions of acollapsible blood container 102 are provided to contain about 220 to 380ml of packed red blood cells, which is equivalent to a unit of packedred blood cells. The height of an oxygen depletion device 10 is furtherconstrained by the presence of one or more sorbent packets and devicesincluded to maintain an appropriate headspace. In view of theseconsiderations, it become apparent that constraints on the dimension ofthe outer receptacle 101 of an oxygen depletion device 10 necessarilylimits the dimensions of a collapsible blood container 102. Accordingly,a collapsible blood container 102 may be divided into one or morechambers in fluid communication with each other.

In aspects according to the present disclosure, an oxygen depletiondevice 10 is designed to be incorporated into existing blood agitationequipment. In certain aspects, an oxygen depletion device 10 isdimensioned to efficiently utilize the space available in agitator andmixing tables. In an aspect, an oxygen depletion device 10 isdimensioned to maximally utilize the area available in a plateletagitator, for example a Helmer Labs Platelet Agitator, Model PF96.Suitable dimensions of an oxygen depletion device 10 include those thatallow for 1, 2, 4, 6, 8, 10 or more bags to be placed on a flat agitatoror mixer surface.

The area of a collapsible blood container 102, within an oxygendepletion device 10 has an area of between about 900 to 1800 cm².Accordingly, an oxygen depletion device 10 that further comprises aspacer 110 effectively doubles the surface area available for gasexchange. In the absence of a spacer 110, the exchange rate of membrane113 of the collapsible blood container 102 on the lower surface issignificantly reduced and the permeable membrane is contacted by theimpermeable film.

The present disclosure provides for, and includes, collapsible bloodcontainers 102 further comprising a tie layer 105, for example asillustrated in FIGS. 1A, 1C, 6, 7, 9A, 9B, 10 and 11. As used herein atie layer 105 comprises an intermediate material that bonds (joins) themembranes 113 (114) together. In certain aspects, the tie layer 105comprises a solid material having a defined shape. As discussed below,tie layers having a defined shape provide for the incorporation ofgeometric features 121 including rounded corners and other mixingenhancing shapes. In certain aspects, a tie layer 105 comprises a liquidor gel that can be dried or cured to provide a joining bond between themembranes 113. Accordingly, a collapsible blood container 102 comprisinga silicone membrane 113 can be joined by a liquid silicone tie layer105. In certain aspects, the silicone rubber tie layer 116 can be liquidsilicone rubber (LSR).

The present disclosure provides for, and includes, collapsible bloodcontainers 102 further comprising a tie layer 105 prepared from a solidmaterial that has a lower melting point than the membranes 113. Byproviding a tie layer 105 having a lower melting temperature, themembranes 113 can be heat joined via the tie layer 105 without damagingthe structure of the microporous membranes, including melting and/orcrystallization. In an aspect, the tie layer 105 is selected to have amelting temperature at least 3° C. below the melting temperature of themicroporous membranes 113. In other aspect, the tie layer 105 has amelting temperature at least 10° C. below the melting temperature of themicroporous membranes 113. In other aspects, a suitable tie layer 105 isselected to maximize the difference in temperature between the tie layerand the microporous membranes 113 (114) to be joined.

In aspects according to the present disclosure, the tie layer 105 isselected from LDPE and the microporous membrane 113 is selected from thegroup consisting of polysulfone, hydrophobic polyvinylidene fluoride(PVDF), cellulose ester, mixed esters of cellulose (MCE),polyethersulfone (PES), polypropylene rendered hydrophobic, andpolyacrylonitrile. In an aspect, the tie layer 105 is LDPE and themicroporous membrane 113 is polysulfone or hydrophobic polyvinylidenefluoride (PVDF). The present disclosure provides for and includes theselection of suitable microporous membranes as provided herein, andfurther includes multilayerd membranes 113 as provided herein.

The present disclosure provides for and includes, construction ofcollapsible blood containers 102 having a tie layer wherein the tielayer extends beyond the seal indicated as gap 109, for example asillustrated in FIG. 9B.

The present disclosure provides for and includes a gap 109 of spacebetween where the seal ends and the tie layer ends. In certain aspects,gap 109 is between 0.05 and 2.5 cm. In other aspects, gap 109 is atleast 0.1 cm wide. In other aspects, gap 109 is at least 0.5 cm wide. Inother aspects, gap 109 is at least 1 cm wide. In other aspects, gap 109is at least 1.5 cm wide. In some aspects, gap 109 is between 0.5 and 1.5cm wide. In other aspects, gap 109 is at least 2 cm wide. In otheraspects, gap 109 is between 2 and 2.5 cm wide. In other aspects, gap 109is at least 2.5 cm wide.

As shown in FIG. 9B, seals 107 are laminated to the membranes 113 andare in turn laminated to each other as seal 108. As illustrated in FIG.7, the lamination of the tie layer 105 can be accomplished in two steps,first to the separate membranes 113, then a second step to join theprelaminated membranes 113 together. In the alternative, the laminationsteps can be combined into a single step wherein a single tie layer 105is used to join membranes together.

As shown in FIG. 9B, seal 107 may extend beyond the width of seal 108.By extending seal 107 beyond the width of seal 108, seal 107 providesfor strengthening of flexure point 115, as indicated in FIG. 9A. Withoutbeing limited to a specific mechanism, it is believed that the tie layer105 acts as a reinforcing strain relief inboard of the seal and allowsfor flexure of the bag at the seal as it is filled and drained of bloodproduct.

The present disclosure provides for, and includes, collapsible bloodcontainers 102 having geometric features that improve the mixing ofblood during the deoxygenation process. The improved geometries of thepresent disclosure further include geometries to enhance the filling anddraining of the collapsible blood containers 102. Improved geometriesreduce or eliminate ‘dead’ spots in the bag. Not to be limited bytheory, dead spot arise in the corners of bags with square geometries.Prior to the present disclosure, methods and blood depletion deviceswere not time limited and the gas exchange methods typically employedresulted in sufficient mixing. Accordingly, the deficiencies of earlierdesigns were not revealed.

In aspects according to the present disclosure, a collapsible bloodcontainers 102 includes one or more geometric features 121. In anaspect, the geometric features comprise rounded corners in thecollapsible blood container 102 and provide for the elimination of‘dead’ spots during mixing. The present disclosure provides for thegeometric features 121 to be incorporated directly into a tie layer 105.In other aspects, the geometric features 121 can be incorporated intothe collapsible blood container 102 using an external die or plate. Inother aspects the geometric features of the collapsible blood container102 can be provided by a suitable mold having the shape of the geometricfeature 121. In certain aspects, the geometric feature 121 provides around or oval shape to a collapsible blood container 102, for example asshown in FIG. 10.

In certain aspects, the geometric feature 121 can be an ellipse with afirst radius from about 0.1 cm to about 7.6 cm and a second radius fromabout 1 cm to about 7.6 cm. In an aspect, the geometric feature 121 canbe an ellipse with a first radius of about 2.5 cm and a second radius ofabout 5 cm. In an aspect, the geometric feature 121 can be an ellipsewith a first radius of about 5 cm and a second radius of about 7.6 cm.In an aspect the geometric feature 121 can be a circle with a diameterof about 5 cm. In an aspect the geometric feature 121 can be a circlewith a diameter of about 7.6 cm.

As is evident, an oxygen depletion device 10 having a defined sizenecessarily constrains the dimensions of a collapsible blood container102 according to the present disclosure. In certain aspects, acollapsible blood container 102 is further limited by a specifiedsurface to volume ratio. In accordance with these limitations, thepresent disclosure provides for, and includes, a collapsible bloodcontainer 102 having two or more chambers in fluid communication witheach other.

The oxygen depletion container device can be constructed in such amanner that allows for the blood volume to area of bag to be optimizedagainst the overall size of the oxygen depletion container device, whileexposing more of the blood volume to the material with oxygenpermeability in the utilized space. The blood volume can be contained ina collapsible blood container 102 having two or more chambers that allowfor their specific arrangement within the outer receptacle 101. Incertain aspects, the oxygen depletion device 10 height, when placed ontoa surface, does not occupy impractical space in the intended mixingapparatus. The chambers can be arranged side to side, stacked on top ofone another, partially stacked onto each other, staggered in a row, orsaddled on top of each other onto one or more stacking heights. Sorbent103 can be positioned over or between chambers as needed. Chambers maybe filled and drained individually or in unison when such chambers areconnected via tubing or fluid conduits that allow for easy filling anddraining. It would be understood that the arrangement andinterconnection of collapsible blood containers 102 having two or morechambers can be performed by a person of skill in the art.

In certain aspects, a collapsible blood container 102 comprises two ormore chambers. In an aspect, a collapsible blood container 102 can havetwo chambers placed side by side or end to end depending on thedimensions. In another aspect, a collapsible blood container 102 canhave three chambers placed side by side or end to end depending on thedimensions. In yet another aspect, a collapsible blood container 102 canhave three chambers placed side by side or end to end depending on thedimensions. A person of ordinary skill could prepare additionalconfigurations of a collapsible blood container 102 having multiplechambers placed in adjacent positions and orientations to maximize theutilization of space.

In other aspects provided for and included in the present disclosure, acollapsible blood container 102 may comprise two or more chambers thatare stacked. When in a stacked configuration, to maintain optimal gasdiffusion rates, spacers 110 or meshes 110 are included to ensure theseparation of adjacent chambers. In certain aspects, the space between astacked chamber further includes one or more sorbent sachets in order tomaintain optimal gas diffusion rates. In certain aspects, two chambersmay be stacked. In another aspect, three chambers may be stacked. In yetanother aspect, four chambers may be stacked.

The present disclosure provides for, and includes, a collapsible bloodcontainer 102 comprising a combination of stacked and adjacent chambers.As provided herein, the number and stacking of chambers of a collapsibleblood container 102 further comprises a surface to volume ratio of thecombined chambers of at least 0.4 cm²/ml. Additional variationsconsistent with the present disclosure can be prepared by one ofordinary skill in the art.

The present disclosure provides for, and includes, an oxygen depletiondevice 10 for depleting oxygen from blood comprising an outer receptacle101 substantially impermeable to oxygen, inner collapsible bloodcontainer 102 that is permeable to oxygen and an oxygen sorbent situatedwithin said outer receptacle wherein the collapsible blood container 102further comprises one or more mixing structures 119 that increase mixingof the blood during oxygen depletion. In certain aspects, the mixingstructures 119 are incorporated into the structure of the collapsibleblood container 102. In other aspects, the mixing structures 119 areadded to the inside of, but not physically joined to the collapsibleblood container 102. In yet other aspects, a mixing structure 119 is astructure outside of the collapsible blood container 102 that restrictsor modifies the shape of the container 102 to decrease or disruptlaminar flow. Mixing structures 119 according to the present disclosureare designed to increase blood movement in the collapsible bloodcontainer 102, increase turbulent flow within the collapsible bloodcontainer 102, or combinations of both. Importantly, mixing structuresand mixing should not significantly increase lysis, or damage to, thered blood cells.

In aspects according to the present disclosure, a mixing structure 119is included in the structure of membrane 113. In certain aspects, amixing structure 119 in membrane 113 comprises ridges, bumps, orprotrusions on the inside of the collapsible blood container 102 and arein contact with the blood. In an aspect, a mixing structure 119 inmembrane 113 comprises one or more ridges. In an aspect, a mixingstructure 119 comprises joining the upper and lower membranes 113 (114)together, for example as illustrated in FIGS. 10C and 10D. In an aspect,the one or more ridges extend across the full width or length of theinner surface of collapsible blood container 102. In other aspects, theridges alternate and may be staggered. In certain aspects, the mixingstructure 119 in membrane 113 comprises bumps or other protrusionsdesigned to disrupt laminar flow and induce turbulence. Similarly, incertain aspects, the mixing structure 119 in membrane 113 comprisesdepressions designed to disrupt laminar flow and induce turbulence. Incertain aspects, the mixing structures 119 are baffles incorporated intomembrane 113. Baffles are flow directing vanes or panels. In someaspects, a mixing structure 119 comprising one or more baffles may beincorporated into a second membrane 114.

In certain aspects, a mixing structure 119 is contained within thecollapsible blood container 102. In an aspect, a mixing structure 119within the collapsible blood container 102 comprises one or more beadsor balls that aid in mixing when the collapsible blood container 102 isagitated. In another aspect, a mixing structure 119 within thecollapsible blood container 102 comprises one or more strings orelongated structures that aid in mixing when the collapsible bloodcontainer 102 is agitated. In yet another aspect, a mixing structure 119within the collapsible blood container 102 comprises a mesh that aids inmixing when the collapsible blood container 102 is agitated.

The present disclosure provides for, and includes, an oxygen depletiondevice 10 having an outer receptacle 101 that is substantiallyimpermeable to oxygen enclosing an inner collapsible blood container 102and providing a headspace. In an aspect, the oxygen sorbent 103 isdisposed within the headspace thereby creating an oxygen depleted statewithin the headspace. In an aspect, said oxygen sorbent 103 disposed inthe headspace further maintains the headspace in an oxygen depletedstate by removing oxygen that may enter through the outer receptacle 101or through the one or more inlets/outlets 30.

Maintaining the headspace in an oxygen depleted state provides forimproved shelf life for oxygen depletion device 10. In an aspect, anassembled oxygen depletion device 10 has a shelf life of at least 24months. In another aspect, the oxygen depletion device 10 has a shelflife of at least 12 months after assembly of the components. In anaspect according to the present disclosure, the assembled oxygendepletion device 10 meets ISTA-2A standards.

In certain aspects of the present disclosure, the headspace provides forimproved processing times. For oxygen depletion device 10, removingambient air present or inert flushing gas from the assembly prior tosealing the outer receptacle 101 reduces the volume of the headspace.Applying a vacuum to the outer receptacle 101 prior to sealing reducesthe volume of the headspace and decreases the total volume of theassembled oxygen depletion device. While reduced overall headspacevolume provides for reduced shipping volume, it can result in increasedfilling times by constraining the collapsible blood container 102. Incertain aspects, the headspace may be flushed with nitrogen gas and thensealed under slightly less than ambient pressure to provide a reducedheadspace volume in the oxygen depletion device 10 without significantlyincreasing the fill and process time.

In certain aspects, the headspace may be initially depleted of oxygen byflushing the headspace with nitrogen. In an aspect, the headspace ofoxygen depletion device 10 is flushed with nitrogen gas prior to sealingthe outer receptacle 101. In an aspect, the flushing gas is >99.9%nitrogen gas.

The present disclosure includes and provides for oxygen depletion device10 having inner collapsible blood container 102 divided into two or morecompartments. In certain aspects, an oxygen depletion device 10, havinga collapsible blood container 102 divided into multiple compartments hasa headspace of between 10 and 500 ml per compartment. In an aspect theheadspace is between 20 and 400 ml per compartment. In another aspectthe headspace volume is between 60 and 300 ml per compartment. In afurther aspect, the headspace volume is between 100 and 200 ml percompartment of a collapsible blood container. In an aspect, an oxygendepletion device 10 having inner collapsible blood container 102 dividedinto compartments has a headspace of about 10 ml per compartment. Inanother aspect, the headspace is about 100 ml to about 200 ml percompartment. In another aspect the headspace is about 300 ml to about500 ml per compartment.

The present disclosure includes and provides for oxygen depletion device10 having inner collapsible blood container 102 divided into two or morecompartments. In certain aspects, an oxygen depletion device 10, havinga collapsible blood container 102 divided into two compartments has aheadspace of between 20 and 1000 ml. In an aspect the headspace isbetween 100 and 800 ml. In another aspect the headspace volume isbetween 200 and 700 ml. In a further aspect, the headspace volume isbetween 300 and 500 ml for a two compartment collapsible bloodcontainer. In an aspect, an oxygen depletion device 10 having innercollapsible blood container 102 divided into two compartments has aheadspace of about 700 ml. In another aspect, the headspace is about 200ml to about 700 ml. In another aspect the headspace is about 300 ml toabout 500 ml.

The present disclosure includes and provides for oxygen depletion device10 having inner collapsible blood container 102 divided into two or morecompartments. In certain aspects, an oxygen depletion device 10, havinga collapsible blood container 102 divided into three compartments has aheadspace of between 20 and 1000 ml. In an aspect the headspace isbetween 100 and 800 ml. In another aspect the headspace volume isbetween 200 and 700 ml. In a further aspect, the headspace volume isbetween 400 and 600 ml for a three compartment collapsible bloodcontainer. In an aspect, an oxygen depletion device 10 having innercollapsible blood container 102 divided into three compartments has aheadspace of about 800 ml. In another aspect, the headspace is about 200ml to about 700 ml. In another aspect the headspace is about 400 ml toabout 600 ml. In an aspect the headspace is about 7000 ml due to fullexpansion of the headspace area. In another aspect the headspace isbetween 700 and 7000 ml. In another aspect the headspace is between 800and 6000 ml. In another aspect the headspace is between 1000 and 5000ml. In another aspect the headspace is between 2000 and 4000 ml.

The present disclosure includes and provides for an oxygen depletiondevice 10 having an inner collapsible blood container 102 and furtherincluding one or more spacers 110 that ensure the separation of theouter receptacle 101 and the inner collapsible blood container 102. Thespacer 110 provides for the maintenance of the headspace in the oxygendepletion device to ensure efficient diffusion of the oxygen from thesurface of membrane 113 to the sorbent 103. A spacer 110 can be preparedfrom one or more of the materials selected from the group consisting ofa mesh, a molded mat, a woven mat, a non-woven mat, a strand veil, and astrand mat. In certain aspects, the spacer 110 can be integrateddirectly into the collapsible blood container 102 as ribs, dimples, orother raised feature that maintains a separation between the outerreceptacle 101 and the inner collapsible blood container 102. Thepresent specification also includes and provides for a spacer 110 to beintegrated into the outer receptacle 101 as ribs, dimples, or othersuitable raised feature capable of maintaining a separation between theouter receptacle 101 and the inner collapsible blood container 102.Mixing is an important aspect of the present disclosure. In one aspectof the present disclosure, spacer 110 is selected to be flexible so asto not interfere with the flow of the blood product.

The present disclosure includes, and provides for, a spacer 110 havingopen areas, for the free diffusion of gas from the surface of thepermeable membranes 113 and 114. In an aspect, the spacer 110 isprovided as a mesh 110 having open spaces 111. As used herein, the openarea 111 is also referred to as the interstice 111. As provided herein,the interstice 111 may be provided by a regular weave of a mesh 110,such that the interstice 111 is regular and repeating within the spacer110. In other aspects, the interstice 111 may comprise an irregular openarea, for example as provided by a spacer 110 constructed from anon-woven mesh. In an aspect, the interstice 111 has an area of betweenabout 0.5 milimeters² (mm²) and about 100 mm². In a further aspect, theinterstice 111 has an area of between 1 mm² and 10 mm². In other aspectsthe interstice 111 has an opening that is greater than 0.75 mm² peropening. In an aspect the open area or interstitial space of a meshcomprise between 30% to 90% of the total area of a spacer 110. In anaspect the open area or interstitial space of a mesh comprise between50% to 80% of the total area of a spacer 110. In a further aspect, theopen area comprises about 60%. In other aspects, the open area comprisesup to 75% of the total area.

The present disclosure provides for, and includes, inner collapsibleblood containers 102 having a spacer 110 incorporated into the membrane113, the membrane 114, or both. In aspects according to the presentdisclosure, the spacer 110 provides for both the separation of the outerreceptacle 101 and the inner collapsible blood container 102 but alsofor the reinforcement of the permeable membranes. In aspects accordingto the present disclosure, the spacer 110 prevents tearing, puncturingand bursting of the inner collapsible blood container 102 when filledwith blood and used in the depletion methods of the present disclosure.In some aspects, the spacer 110 is provided as a mesh 110 that isintegrated into a silicone membrane during the manufacturing process. Inother aspects, the spacer 110 is applied to, and joined to, a finishedsilicone membrane. In other aspects, the spacer 110 is provided as anintegrated mesh of a porous membrane.

In an aspect, a membrane 113 or 114 having an integrated spacer 110 isprepared from a suspension of liquid silicone rubber (LSR). In anaspect, the LSR is suspended in xylene, hexane, tert butyl acetate,heptane, acetone, or naptha. In aspects according to the presentdisclosure, the suspension comprises 10 to 30% LSR. As provided herein,a membrane 113 or 114 having an integrated spacer 110 is prepared byproviding a 20 to 750 μm layer of an LSR suspension, partially curingthe LSR layer and applying a spacer 110 as provided in the presentdisclosure and performing a second curing step to provide a cured 10 to100 μm thick silicone membrane 113 having an integrated spacer 110.

The present disclosure also includes and provides for a mesh 110comprising co-extruded fibers having an inner material 117 and bindingmaterial 118. In aspects according to the present disclosure, bindingmaterial 118 is integrated into the pores of membrane 113 (114) duringapplication of the mesh 110 to the membrane. In an aspect, the bindingmaterial 118 is integrated into the pores of a porous membrane 113 byheating. In aspects according to the present disclosure, bindingmaterial 118 may be selected from the group consisting of ethylvinylalcohol (EVOH), ethylvinylacetate (EVA), or acrylate. In aspectsaccording to the present disclosure co-extruded fibers having an innermaterial 117 and binding material 118 are meshes 110 that include theDuPont Bynel® series of modified ethyl vinyl acetates and modified ethylvinyl acrylates.

The present disclosure also includes and provides for inner collapsibleblood containers 102 that further comprise a window 112. As used herein,a window 112 is made of a transparent material and is bonded orotherwise incorporated into the inner collapsible blood container 102.In accordance with the present disclosure, suitable materials for window112 are blood compatible. In certain aspects, materials suitable for awindow 112 are oxygen impermeable. In other aspects, materials suitablefor a window 112 are oxygen impermeable. The size of a window 112 needonly be large enough to provide observation of the blood.

Also included and provided for by the present disclosure are collapsibleblood containers having bis(2-ethylhexyl) phthalate (DEHP). DEHP isincluded in most PVC based blood storage bags as a plasticizer where ithas been observed that DEHP provides a protective effect to stored redblood cells. See U.S. Pat. No. 4,386,069 issued to Estep. In certainaspects, an oxygen depletion device 10 may further include DEHPincorporated in the inner collapsible blood container 102. In otheraspects, DEHP may be provided separately within the inner collapsibleblood container 102.

The present disclosure provides for, and includes, an oxygen depletiondevice 10 that does not include DEHP. It has been hypothesized that DEHPmay act as an endocrine disruptor and certain regulatory agencies areconsidering ordering the removal of DEHP from blood bags. It has beenobserved that DEHP may not be necessary when red blood cells are storedanaerobically. See, International Patent Publication No. WO 2014/134503,hereby incorporated by reference in its entirety. Accordingly, incertain aspects, oxygen depletion device 10 entirely excludes DEHP fromall blood contacting surfaces. In other aspects, oxygen depletion device10 limits DEHP containing surfaces to tubing, ports, and inlets such asthose illustrated in the Figures at, for example, 106 and 205. In anaspect, oxygen depletion device 10 excludes a DEHP containingcollapsible blood container 102.

The present disclosure provides for, and includes, an oxygen depletiondevice 10 having an oxygen indicator 104. Similarly, the presentdisclosure provides for, and includes, blood storage device 20 having anoxygen indicator 206. In an aspect, the oxygen indicator 206 detectsoxygen and indicates that the oxygen depletion device 10 has beencompromised and is no longer suitable for its intended purpose. In anaspect, the oxygen indicator 206 provides a visual indication of thepresence of oxygen. In certain aspects, the oxygen indicator 206provides an indication of the amount of oxygen.

In an aspect according to the present disclosure, the outer receptaclemay contain an oxygen indicator to notify the user if the oxygen sorbentis no longer active for any reason, such as age, or if the outerreceptacle has been compromised, allowing excess oxygen to ingress fromambient air. Such oxygen indicators are readily available and are basedon a methylene blue indicator dye that turns blue in the presence ofoxygen of about 0.5% or more and pink when the oxygen level is belowabout 0.1%. Examples of these oxygen indicators are the Tell-Tab oxygenindicating tablet from Sorbent Systems, Inc. (Impak Corp., Los Angeles,Calif.), and the Oxygen Indicator tablet from Mitsubishi Gas ChemicalAmerica (MGCA, NY, N.Y.).

The present disclosure provides for, and includes, methods for preparingblood for storage under oxygen depleted conditions comprising providingblood having red blood cells having oxygen to be removed to an oxygendepletion device 10, incubating the blood for a period of time, andtransferring the deoxygenated blood to an anaerobic storage bag. Inaspects according to the present disclosure, the method further includesagitating the oxygen depletion device 10 to provide for mixing of theblood for deoxygenation. In other aspects, due to the configuration ofthe oxygen depletion device 10, agitating is not necessary.

For safety, the collection and processing of blood is regulated by anational or regional governmental agency. In the U.S., the Food and DrugAdministration (FDA) has established guidelines for the proper handlingof blood and blood products. Similarly in Europe, the European Union hasbeen granted regulatory authority that is binding for member states, andtypically follows guidelines provided by the Council of Europe. The keyrequirements for blood establishments and for hospital blood banks inthe United Kingdom (UK), for example, are defined in the Blood Safetyand Quality Regulations (Statutory Instrument 2005 No. 50) and areenforced by the Medicines and Healthcare products Regulatory Agency,whose powers are derived from UK legislation, to maintain the safety andquality of blood and blood products for transfusion within the UK.

Generally, the guidelines established by the various authorities fall into two main groups. In the first group, as exemplified by the U.S., theallowable time period from donor collection to processing for platelets,and thus driving the storage of RBC's at 2 to 6° C., is 8 hours. Thatis, the various processing steps, currently including plasma separationand collection, leukoreduction, platelet separation and collection andpacked red blood cell preparation, need to be completed, and variouscomponents stored within 8 hours in order to preserve the viability ofthe platelets (see More & Holme, “Concepts about current conditions forthe preparation and storage of platelets” in Transfus Med Rev 1991;5:48-59). In Europe, the period available for processing is 24 hours.Accordingly the methods and processes provided in the present disclosureare designed to achieve the beneficial, and storage lesion reducing,level of deoxygenation for blood storage within about 8 hours fromvenipuncture.

In accordance with the methods of the present disclosure, blood may beobtained from a donor and processed to less than 20% oxygen saturationwithin 12 hours of collection. Beginning the depletion process, at orsoon after collection, improves the efficiency of the process byleveraging the increases in reaction rates due to the highertemperatures. In an aspect, the blood is collected from a donor at about37° C. and collected in an oxygen depletion device 10 having a suitableamount of anticoagulant. In addition to the increased temperature, thewhole blood is typically about 35-65% oxygen saturated when collected byvenipuncture from a patient. In one aspect of the present disclosure thewhole blood is 35-65% oxygen saturated when collected by venipuncturefrom a patient. In another aspect the whole blood is 40-60% oxygensaturated when collected by venipuncture from a patient. In anotheraspect the whole blood is 45-55% oxygen saturated when collected byvenipuncture from a patient. In another aspect the whole blood is 50-65%oxygen saturated when collected by venipuncture from a patient.Conventional methods do not provide collection kits and bags thatprevent the ingress of oxygen. Thus delays in beginning the oxygenreduction process can greatly increase the time necessary to prepareoxygen-reduced blood having less than 20% oxygen saturation.

The methods and devices of the present disclosure further provide forthe preparation of oxygen-reduced blood having less than 10% oxygensaturation. In an aspect, the 10% level is achieved within 8 hours orless of collection from a donor. In other aspects, the blood is reducedto less than 10% oxygen saturation in 6 hours or less. In yet otheraspects, the blood is reduced to less than 10% oxygen saturation in 4hours or less.

As used herein, the term “blood” refers to whole blood, leukoreducedRBCs, platelet reduced RBCs, and leukocyte and platelet reduced RBCs.The term blood further includes packed red blood cells, platelet reducedpacked red blood cells, leukocyte reduced packed red blood cells(LRpRBC), and leukocyte and platelet reduced packed red blood cells. Thetemperature of blood can vary depending on the stage of the collectionprocess, starting at the normal body temperature of 37° C. at the timeand point of collection, but decreasing rapidly to about 30° C. as soonas the blood leaves the patient's body and further thereafter to roomtemperature in about 6 hours when untreated, and ultimately beingrefrigerated at between about 2° C. and 6° C.

As used herein, the term “whole blood” refers to a suspension of bloodcells that contains red blood cells (RBCs), white blood cells (WBCs),platelets suspended in plasma, and includes electrolytes, hormones,vitamins, antibodies, etc. In whole blood, white blood cells arenormally present in the range between 4.5 and 11.0×10⁹ cells/L and thenormal RBC range at sea level is 4.6-6.2×10¹²/L for men and4.2-5.4×10¹²/L for women. The normal hematocrit, or percent packed cellvolume, is about 40-54% for men and about 38-47% for women. The plateletcount is normally 150-450×10⁹/L for both men and women. Whole blood iscollected from a blood donor, and is usually combined with ananticoagulant. Whole blood, when collected is initially at about 37° C.and rapidly cools to about 30° C. during and shortly after collection,but slowly cools to ambient temperature over about 6 hours. Whole bloodmay be processed according to methods of the present disclosure atcollection, beginning at 30-37° C., or at room temperature (typicallyabout 25° C.). As used herein, a “unit” of blood is about 450-500 mlincluding anticoagulant.

As used herein, a “blood donor” refers to a healthy individual from whomwhole blood is collected, usually by phlebotomy or venipuncture, wherethe donated blood is processed and held in a blood bank for later use tobe ultimately used by a recipient different from the donor. A blooddonor may be a subject scheduled for surgery or other treatment that maydonate blood for themselves in a process known as autologous blooddonation. Alternatively and most commonly, blood is donated for use byanother in a process known as heterologous transfusion. The collectionof a whole blood sample drawn from a donor, or in the case of anautologous transfusion from a patient, may be accomplished by techniquesknown in the art, such as through donation or apheresis. Whole bloodobtained from a donor using venipuncture has an oxygen saturationranging from about 30% to about 70% saturated oxygen (sO₂).

As used herein, “red blood cells” (RBCs) includes RBCs present in wholeblood, leukoreduced RBCs, platelet reduced RBCs, and leukocyte andplatelet reduced RBCs. Human red blood cells in vivo are in a dynamicstate. The red blood cells contain hemoglobin, the iron-containingprotein that carries oxygen throughout the body and gives red blood itscolor. The percentage of blood volume composed of red blood cells iscalled the hematocrit. As used herein, unless otherwise limited, RBCsalso includes packed red blood cells (pRBCs). Packed red blood cells areprepared from whole blood using centrifugation techniques commonly knownin the art. As used herein, unless otherwise indicated, the hematocritof pRBCs is about 50%.

Platelets are small cellular components of blood that facilitate theclotting process by sticking to the lining of the blood vessels, andalso facilitate healing by releasing growth factors when activated. Theplatelets, like the red blood cells, are made by the bone marrow andsurvive in the circulatory system for 9 to 10 days before they areremoved by the spleen. Platelets are typically prepared using acentrifuge to separate the platelets from the buffy coat sandwichedbetween the plasma layer and the pellet of red cells.

Plasma is a protein-salt solution and the liquid portion of the blood inwhich red and white blood cells and platelets are suspended. Plasma is90% water and constitutes about 55 percent of the blood volume. One ofthe primary functions of plasma is to assist in blood clotting andimmunity. Plasma is obtained by separating the liquid portion of theblood from the cells. Typically, plasma is separated from the cells bycentrifugation. Centrifugation is the process used to separate thecomponents of the whole blood into the plasma, the white blood cells,the platelets and the packed red blood cells. During centrifugation, theplasma will initially migrate to the top of a vessel during a lightspin. The plasma is then removed from the vessel. The white blood cellsand platelets are removed during a second centrifugation cycle toproduce the packed red blood cells.

The present disclosure includes and provides for methods for preparingoxygen depleted blood for storage. An oxygen reduced blood or bloodcomponent suitable for storage and benefiting from the reduced damagefrom storage lesions, reduced toxicity, and importantly reducedmorbidity, is a blood or blood component having an oxygen saturation ofless than about 20%. In certain aspects, the oxygen levels in the bloodor blood component are reduced to a level of less than 15%. In otheraspects, the oxygen saturation of the blood is reduced to 10% or lessprior to storage. In yet another aspect, the oxygen saturation of theblood is reduced to less than 5% or less than 3% prior to storage.

According to methods of the present disclosure, the blood or bloodcomponent is depleted of oxygen and placed into storage within 4 to 24hours of collection. In other aspects, the methods provide for thedepletion of oxygen and placement into storage within 8 hours ofcollection. In other aspects, the blood or blood component is depletedof oxygen and placed into storage in less than 6 hours of collection. Inyet another aspect, the blood is depleted of oxygen and placed intostorage in less than 4 hours of collection.

The present disclosure provides for, and includes, methods for preparingblood for storage under oxygen depleted conditions comprising providingblood having red blood cells having oxygen to be removed to an oxygendepletion device 10, and incubating the blood for a period of time. Incertain aspects, the blood is mixed through agitation. In other aspects,the oxygen depletion device provides for sufficient deoxygenation withlittle or no mixing.

As would be understood, blood for depletion may start with varyinglevels of oxygen saturation. In certain aspects, the blood is wholeblood collected at about 70% saturation and between about 40% to 45%hematocrit. The methods of the present disclosure also provide for therapid deoxygenation of LRpRBCs that typically have a hematocrit of about50% and saturation levels of up to 90% or higher.

The devices and methods of the present disclosure are intended toprovide oxygen depleted blood for storage within 24 hours or less. Incertain aspects, the oxygen is removed using an oxygen depletion device10 by incubation for a time period with agitation. In other aspects, theoxygen is removed using an oxygen depletion device 10 using methods inwhich the depletion device is not agitated or otherwise mixed during theincubation period. As would be understood by one of skill in the art,inclusion of an agitation or mixing step in the process allows for anoxygen depletion device 10 to have lower surface to volume ratios.Agitation can also reduce the permeability necessary to achieve adesired level of deoxygenation. To achieve the most rapid depletionkinetics, a combination of an oxygen depletion device 10 having highpermeability and a high surface to volume ratio is combined withagitation during the depletion period. Depending on the application andthe processing protocols employed, the time necessary to completeprocessing can vary to between 4 and 24 hours. Thus, the devices andmethods of the present disclosure can be incorporated into the protocolsof existing blood processing centers and comply with applicable regionalregulations by adjusting the devices and methods as provided in thepresent disclosure.

In aspects according to the present disclosure, to reduce the processingtime to achieve a blood saturation of less than 20%, the blood can beagitated or mixed during the depletion period. In most aspects, theblood is agitated or mixed for less than 24 hours. As mixing andagitation of the blood during processing can lead to lysis anddegradation, the depletion time period with agitation should beminimized.

In certain aspects, the blood is incubated with agitation for less than12 hours. In other aspects, the incubation and agitation time is lessthan 8 hours. Also provided for are methods for reducing oxygen to lessthan 20% using an oxygen depletion device 10 and incubating withagitation for less than 6 hours or less than 4 hours. In yet otheraspects, the incubation time with agitation is 3.5 hours or 3.0 hours.In certain aspects, the blood can be reduced to 20% or less with a4-hour incubation with agitation in an oxygen depletion device 10. Inyet further aspects, the method provides for incubation times of 0.5 or1.0 hour. In other aspects, blood is incubated for 1.5 hours or 2.0hours in an oxygen depletion device 10.

It is well understood that reaction rates are temperature dependent,with higher temperatures increasing the reaction rate. The rate constantk varies exponentially with temperature where k=Ae^(−Ea/RT) (theArrhenius equation). Notably, the dependence on temperature isindependent of the concentration of reactants and does not depend onwhether the order of the rate is constant (e.g., first order vs. secondorder). Typically, a 10° C. increase in temperature can result in a twofold increase in reaction rate. Accordingly, a person of skill in theart would recognize that the release of oxygen from hemoglobin as wellas the other steps of the deoxygenation process is temperaturedependent. Importantly, once the temperature of the blood is lowered tothe standard storage temperature of between 2° C. and 6° C., the rate ofdeoxygenation is significantly reduced. Even further, under currentapproved protocols for the collection, processing and storage of bloodfor transfusion purpose conditions, the stored blood is not mixed whichfurther reduces the rate at which oxygen can be removed. Accordingly,the methods and devices of the present disclosure are designed to removemost of the oxygen prior to storage and within the time periodsestablished by the appropriate regulatory agencies. As provided herein,the depletion of oxygen is intended to begin as soon after collectionfrom the donor as possible, and is intended to be largely completedprior to cooling the blood for storage.

As provided herein, the methods can be performed using recentlycollected blood that is about 37° C., when it is collected from thedonor. In other aspects, the blood may be processed prior to depletion,including removing leukocytes, plasma, and platelets. In thealternative, the blood may be further processed after oxygen reduction.

The present disclosure provides for, and includes, processing blood thathas cooled from body temperature to ambient temperature, typically about25° C. Using the methods and devices disclosed here, oxygen reducedblood having less than 20% oxygen saturation can be prepared at ambienttemperatures (e.g., about 25° C.). The ability to reduce the oxygen todesired and beneficial levels at ambient temperatures allows for thesystems and methods to be incorporated into existing blood collectionprotocols and blood collection centers.

The present disclosure provides for, and includes, methods for preparingblood for storage under oxygen depleted conditions comprising providingblood having red blood cells having oxygen to be removed to an oxygendepletion device 10, incubating the blood for a period of time andfurther comprising agitating or mixing during the incubation period. Asused herein, the terms “agitating” or “mixing” are used interchangeablyand include various mixing methods, including but not limited torocking, nutating, rotating, stirring, massaging, swinging,linearly-oscillating and compressing the oxygen depletion device.

In a method according to the present disclosure, the incubation periodwith agitation can be as short as 30 minutes and up to 24 hours. Incertain aspects, the method includes an incubation period of between 1and 3 hours with agitation in an oxygen depletion device 10. In otheraspects, the incubation period is between 1 and 4 hours or 1 and 6hours. In other aspects, the incubation period is about 2 hours or about4 hours.

In an aspect according to the present disclosure, a method of reducingthe oxygen from red blood cells includes placing the red blood cells ina device according to the present disclosure and placing the device onan agitator to enhance the oxygen removal from the red blood cellsthrough a mixing action. The use of agitators in the practice of bloodtransfusion is well known with respect to preventing clot formation,such as in the use of rocker tables and donation scale mixers, whichprovide for a gentle rocking motion of about 7 degrees tilt and 1 toabout 15 oscillations per minute. Similar devices, already available inoxygen depletion centers and familiar to staff, can be used to ensureproper mixing.

To maximize the kinetics of the oxygen depletion process, both physicaland methodological approaches can be applied. As discussed above,physical approaches to reducing the resistance to diffusion of the innerblood compatible bag is achieved by selecting materials with highpermeability and by reducing the thickness of the material to decreasethe Barrer value. For microporous materials, apparent Barrer values canbe decreased by decreasing the size of the micropores and by increasingthe number of micropores. It is understood that the size of microporesare necessarily limited by the necessity to prevent the perfusion ofwater through the barrier which occurs in certain microporous materialsat about 1 μm. Also as provided above, the surface to volume ratio isselected to reduce the diffusion distance of the dissolved oxygen as itmakes its way to the permeable surface. These limitations andrequirements of the materials and design to achieve effective and rapidreduction of oxygen in blood are discussed above.

In addition to minimizing the diffusion barriers and the diffusiondistance through design and by appropriate selection of materials, theeffective diffusion distance can be further reduced by appropriatemixing. As would be understood, complete and efficient mixingeffectively eliminates the effect of diffusion distance on the bloodreduction process as oxygen containing red blood cells enter the oxygenfree environment in close proximity to the permeable membrane.Similarly, the diffusion distance would also be eliminated by spreadingthe blood into an impracticably thin volume. The present disclosureprovides methods and devices that optimize the devices and methods toachieve high rates of depletion.

The present disclosure provides for, and includes, methods of mixingblood in an oxygen depletion device 10 that achieves rapid rates ofdeoxygenation and rate constant of between about 0.5×10⁻² min⁻¹ andabout 5.0×10⁻² min⁻¹. In aspects according to the present disclosure,the rate constant is at least −1.28×10⁻² min⁻¹. In other aspects,deoxygenation occurs at rate having a rate constant of at least−0.5×10⁻². In another aspect, deoxygenation occurs at rate having a rateconstant of at least −0.9×10⁻². In another aspect, deoxygenation occursat rate having a rate constant of at least −1.0×10⁻². In another aspect,deoxygenation occurs at rate having a rate constant of at least−1.5×10⁻². In further aspects, deoxygenation occurs at rate having arate constant between −1.0×10⁻² min⁻¹ and −3.0×10⁻² min⁻¹. In furtheraspects, deoxygenation occurs at rate having a rate constant between−1.0×10⁻² min⁻¹ and −2.0×10⁻²min⁻¹. In further aspects, deoxygenationoccurs at rate having a rate constant between −1.0×10⁻² min⁻¹ and−4.0×10⁻² min⁻¹.

In an aspect according to the present disclosure, proper mixing isachieved in an oxygen depletion device 10 having a surface to volumeratio of at least 5.0 cm²/ml. Not to be limited by theory, it ishypothesized that at lower surface to volume ratios, the collapsibleblood container does not have sufficient capacity to allow for themovement of blood and no mixing occurs. It would be appreciated that abag, filled to capacity like an engorged tick, would be essentiallyrefractory to mixing and no convection or other currents could bereadily induced. In other words, an inner collapsible container 102 thatis filled to capacity to the extent that the flexibility of the bagmaterial is reduced beyond its ability to yield during agitation resultsin essentially no mixing. Accordingly, by selecting a surface to volumeratio of at least 4.85 cm²/ml mixing can occur as the blood is ‘sloshed’around. It would be appreciated that improper mixing leads toundesirable hemolysis of the red blood cells. Accordingly, mixing alsohas practical limits. The present disclosure provides for devices andmethods to reduce potential hemolysis while achieving significantmixing.

In an aspect according to the present disclosure, a method of reducingthe oxygen from red blood cells includes placing the red blood cells ina device according to the present disclosure and placing the device onan agitator to enhance the oxygen removal from the red blood cells. Theuse of agitators in the practice of blood transfusion is well known withrespect to preventing clot formation, such as in the use of rockertables and donation scale mixers when used with whole blood andsuspensions of red blood cells, and also for platelet storage, whereinthe platelets require oxygen for survival and agitation to preventclumping and activation of the platelets.

With respect to currently available devices for agitating red bloodcells, whether whole blood or other red blood cell suspensions, aplatform is typically rotated a few degrees about a central axis toprovide for a gentle rocking motion and there are many available choicescommercially available. For example, Bellco Glass model #7740-10000(Bellco Glass, Inc., Vineland, N.J.) provides for 7 degrees tilt and 1to about 12 oscillations per minute. The Medicus Health model 5277M5nutating mixer (Medicus Health, Kentwood, Mich.) provides for a 20degree angle of inclination at 24 rpm for the suspension of red bloodcell samples, while another style device used at the time of donation toprevent clotting of whole blood is the Genesis blood collection mixermodel CM735A (GenesisBPS, Ramsey, N.J.), which provides for about 20degrees of tilt and performs 3 cycles in about 3 seconds, then rests forabout 2 seconds to weigh the sample and repeats until the desired weightis achieved. The Benchmark Scientific model B3D2300 (BenchmarkScientific, Inc., Edison, N.J.) provides for a variable 0-30 degree tiltangle and 2-30 oscillations per minute.

The present disclosure further includes and provides for other availablemeans of agitation of blood samples including orbital shakers, such asthe model LOS-101 from Labocon (Labocon Systems, Ltd, Hampshire, U.K.),having a displacement of 20 mm and an oscillation rate of 20-240 rpm, orthe model EW-51820-40 from Cole-Parmer (Cole-Parmer, Inc., Vernon Hills,Ill.) having a displacement of 20 mm and an oscillation rate of 50-250rpm.

Devices to agitate platelets are also well known in the art and includevarious models such as the PF96h from Helmer Scientific (HelmerScientific, Noblesville, Ind.) which provides for a linear oscillationof about 70 cycles per minute with a displacement of about 38 mm (1.5inches), and the model PAI 200 from Terumo Penpol (Terumo Penpol Ltd.,Thiruvananthapuram, India) with an oscillation of about 60 cycles perminute and a displacement of about 36 mm (1.4 inches).

While the devices of the present disclosure provide for enhanceddeoxygenation of red blood cells, the use of modified motions providesfor even further oxygen removal from the red blood cells. It is wellknown that platelets can be activated by mechanical agitation, such asshear force, and are therefore subject to limitations on how muchphysical agitation can be tolerated before such activation occurs.Hemolysis of red blood cells is estimated to occur at shear stresslevels above approximately 6000 dyne/cm² (Grigioni et al., J. Biomech.,32:1107-1112 (1999); Sutera et al., Biophys. J., 15:1-10 (1975)) whichis an order of magnitude higher than that required for plateletactivation (Ramstack et al., J. Biomech., 12:113-125 (1979)). In certainaspects, currently available platelet agitators operating at about 36 mmdisplacement and about 65 cycles per minute (cpm) provide fordeoxygenation of red blood cells as disclosed herein. In other aspects,improved rates and extent of deoxygenation without hemolysis areachieved using linear oscillating motion with a displacement of between30 mm and about 125 mm. In another aspect, the agitation is a linearoscillation from about 50 mm to about 90 mm.

The present disclosure also provides for, and includes, adjusting thefrequency of oscillation to ensure efficient mixing. In addition toplatelet shakers having a displacement of about 36 mm and a frequency ofabout 65 cpm, in certain aspects, the frequency is from about 60 toabout 150 cycles per minute (cpm). In certain aspects, the frequency ofagitation is between about 80 and about 120 cpm.

In certain aspects according to the present disclosure, when using anagitator or mixer, various configurations of the chambers in acollection device 10 with more than one chamber are provided. With anagitator that moves in a horizontal motion, in one aspect, two to eighthorizontal (flat on surface) chambers are arranged, side by side, end toend, one on top of the other, one on top of the other with one or morepartially covering the chamber(s) beneath. In other aspects, with anagitator that moves in a vertical motion, two to eight vertical(perpendicular to surface) chambers are arranged, side by side, end toend, one on top of the other, one on top of the other with one or morepartially covering the chamber(s) beneath. In another aspect, with anagitator that moves up and down traversing an angle >0 and <90 degreesto horizontal two to eight upright (>0 degrees <90 degrees tohorizontal) chambers are arranged, side by side, end to end, one on topof the other, one on top of the other with one or more partiallycovering the chamber(s) beneath.

A further advantage of the agitation and mixing of the oxygen depletiondevice 10 is that the movement of the blood or blood component caused bythe agitator also moves the sorbent sachet located on the top or bottomof the oxygen depletion device 10. Because the sorbent sachet is movingup and down as it rests on top, the active ingredient that absorbs theoxygen in the headspace is constantly settling. This constant movementof the active ingredient moves oxidized iron particles out of the way ofnon-oxidized iron particles, speeding up the oxygen absorption potentialof the sorbent.

The present disclosure provides for, and includes, mixing the oxygendepletion device 10 by compression of the collapsible blood container102. Compressing of the collapsible container 102 is achieved byapplying pressure on one of the collapsible container's larger surfacesat 30 cm/sec during 1-3 seconds creating a hydrostatic pressure of100-300 mmHg within the collapsible container, then applying a pressureto the opposite surface of the collapsible container at 10-30 cm/secduring 1-3 seconds creating a hydrostatic pressure of 100 to 300 mmHgwithin the collapsible container. This operation is to be performed for2 to 4 hours.

The present disclosure provides for, and includes, mixing the oxygendepletion device 10 by massage of the collapsible blood container 102.Massaging of the collapsible container 102 is achieved by displacing aroller type device along one surface of the collapsible containercompleting the full translation in 1-3 seconds and making thecollapsible container collapse and agitate its contents. This operationis to be performed for 1-2 hours. In another aspect, a roller travelsalong another surface of the collapsible container, completing the fulltranslation in 1-3 seconds and making the collapsible container collapseand agitate its content. This operation is to be performed for 1-2hours.

The present disclosure provides for, and includes, a blood storagedevice 20, for storing oxygen depleted blood and maintaining the bloodin a deoxygenated state during the storage period. Certain anaerobicblood storage devices (ASB) are known in the art, including for exampleU.S. Pat. No. 6,162,396 to Bitensky et al. The anaerobic blood storagedevices of the prior art did not include ports and inlets designed to besubstantially impermeable to oxygen. Accordingly, the prior artanaerobic storage devices had poor shelf lives prior to use and weresusceptible to significant ingress of oxygen. As provided in the presentdisclosure, an improved blood storage device 20 comprising featuresdirected to maintaining the integrity of the device while allowing forthe sampling of the blood that occurs during storage and blood banking.The improved ASB also provides for improved diffusion of oxygen from theblood, providing for additional depletion during the storage period.

The blood storage device 20 comprises an outer receptacle 201 that issubstantially impermeable to oxygen, a collapsible blood container 202comprising a locating feature 203 adapted to align the collapsible bloodcontainer 202 within the geometry of the outer receptacle 201; at leastone inlet/outlet 30 comprising connecting to the collapsible bloodcontainer 202 and a bond 302 to the outer receptacle 201, wherein thebond 302 to the outer receptacle 201 is substantially impermeable tooxygen and an oxygen sorbent 207 situated within the outer receptacle201.

As used herein, an outer receptacle 201 is at least equivalent to anouter receptacle 101. Also as used herein, an inner collapsible bloodcontainer 202 includes blood containers as provided above for an innercollapsible blood container 102 but also provides for collapsible bloodcontainers 202 comprising materials that are less permeable to oxygen,such as PVC. Also as provided herein, oxygen sorbent 207 is at leastequivalent to sorbent 103 and may be provided in sachets as discussedabove.

Also included and provided for in the present disclosure are bloodcollecting kits. In an aspect according to the present disclosure, anoxygen depletion device for depleting oxygen from blood is included in ablood collection kit that reduces or eliminates the introduction ofoxygen during the blood collection process. Blood collection kits in theart do not include any features or elements that prevent theintroduction of oxygen during the collection process. Accordingly, kitsin the art having multiple containers provide from about 3 cc's ofresidual oxygen per container, plus additional ingress through materialsand fittings, and thereby increase the saturation of oxygen (sO2) fromthe venous oxygen saturation (SvO2) of about 40 to 60% to up to fullsaturation. In an aspect according to the present disclosure, the entireblood collection kit is contained in an oxygen free or oxygen reducedenvironment. In an aspect, the blood collection kit is contained withina kit enclosure bag that is substantially oxygen impermeable andincludes within the enclosure bag an amount of oxygen sorbent absorboxygen. Amounts of sorbent for a blood collection kit according to thepresent disclosure are separate from, and in addition to amounts ofsorbent that may be included in a blood collection bag or anaerobicstorage bag.

In certain aspects according to the present disclosure, the amount ofoxygen sorbent included in a blood collection kit is sufficient toremove oxygen from a blood collection kit introduced during manufacture.In an aspect, the blood collection kit includes oxygen sorbentsufficient to absorb 10 cc's of oxygen. In another aspect, the bloodcollection kit includes oxygen sorbent sufficient to absorb 60 cc's ofoxygen. In another aspect, the blood collection kit includes oxygensorbent sufficient to absorb 100 cc's of oxygen. In another aspect, theblood collection kit includes oxygen sorbent sufficient to absorb 200cc's of oxygen. In another aspect, the blood collection kit includesoxygen sorbent sufficient to absorb 500 cc's of oxygen. In anotheraspect, the blood collection kit includes oxygen sorbent sufficient toabsorb from 10 to 500 cc's of oxygen. In another aspect, the bloodcollection kit includes oxygen sorbent sufficient to absorb up to 24,000cc to allow for the management of shelf life of the device. In certainaspects according to the present disclosure, the oxygen sorbent isdisposed in one or more sachets.

In an aspect, the amount of oxygen sorbent is sufficient to maintain anoxygen depleted environment for the blood collection kit during thestorage. In certain aspects, oxygen is flushed from the blood collectionkit during manufacture. Accordingly, the amount of oxygen sorbent may bereduced to account for leakages and residual permeability of thesubstantially impermeable kit enclosure bag.

Also included and provided for in the present disclosure are additivesolution bags that are substantially impermeable to oxygen. In an aspectaccording to the present disclosure, substantially oxygen impermeableadditive solution bags avoid the reintroduction of oxygen to the oxygenreduced blood after oxygen reduction in the oxygen reduction bloodcollection bag.

In aspects of the present disclosure, the method may further includeadding an additive solution to the packed RBCs to form a suspension. Incertain aspects, the additive solution may be selected from the groupconsisting of AS-1, AS-3 (Nutricel®), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP,SOLX, ESOL, EAS61, OFAS1, and OFAS3, alone or in combination. AdditiveAS-1 is disclosed in Heaton et al., “Use of Adsol preservation solutionfor prolonged storage of low viscosity AS-1 red blood cells,” Br JHaematol., 57(3):467-78 (1984). In a further aspect, the additivesolution may have a pH of from 5.0 to 9.0. In another aspect, theadditive may include an antioxidant. In some aspects according thepresent disclosure, the antioxidant may be quercetin, alpha-tocopheral,ascorbic acid, or enzyme inhibitors for oxidases.

EXAMPLES Example 1 Fabrication of Outer Receptacle 101

A barrier bag is fabricated by heat sealing along one edge by placing apair of RollPrint Clearfoil® Z film #37-1275 (Rollprint PackagingProducts, Inc., Addison, Ill.) sheets about 23×30.5 cm (9×12 inches)into a heat sealer along the shorter 23 cm length. A piece of multilayertubing having a polyethylene outer layer, a PVC inner layer, and anintermediary bonding layer of EVA (Pexco, Inc., Athol, Mass. orExtrusion Alternatives, Inc., Portsmouth, N.H.) 0.4 cm I.D. by 0.55 cmO.D. by about 2.6 cm long is placed onto a solid brass mandrel about 0.4cm diameter by about 2.5 cm length and then placed between the films andlocated in the transverse groove of the heat sealing dies heated toabout 130° C. The press is activated and set to about 4 seconds durationat 21×10⁴ Pascal (Pa) to create a continuous welded seal along thelength of the dies, with the short piece of multilayer tubing sealed inplace. The short multilayer tubing provides for an oxygen impermeableseal around the outer diameter of the tubing while also providing fluidconnectivity through the seal. A piece of PVC tubing 0.3 cm I.D.×0.41 cmO.D. by about 30.5 cm length (Pexco, Inc., Athol, Mass., or ExtrusionAlternatives, Inc., Portsmouth, N.H.) is solvent bonded usingcyclohexanone to the multilayer tubing from the outside of the bag.

Sealing of the two long edges of the barrier film is performed with animpulse heat sealer (McMaster Carr #2054T35, McMaster Carr, Inc.,Robbinsville, N.J.), leaving the last remaining short edge of thebarrier bag unsealed to place a blood container 102 inside.

Example 2 Preparation of Silicone Sheets

Liquid Silicone Rubber (LSR)

Silicone sheets having a thickness of about 25 μm are fabricated bymixing equal parts of a two-part silicone elastomer dispersion in asuitable solvent, such as xylene, for example NuSil MED10-6640. MED10-6640 is supplied as a 2 part resin system. As the first step, Part Aand Part B are mixed in equal measure to create the dispersion. Next,the air was removed under vacuum. The vacuum time was selected to ensurethat no bubbles were left in the dispersion. Next the dispersion isspread out and passed under a precision knife edge on a custom builtknife coating tray. The sheet is partially cured by heating beforeplacing a sheet of polyester mesh fabric (Surgical Mesh, Inc.,Brookfield, CT #PETKM3002) onto the partially cured silicone sheet. Thepolyester mesh fabric is pressed into the partially cured sheet byapplying a load onto the laminate. The laminate is cured using a rampcure using the following sequence of time and temperature combination:30 minutes at ambient temperature and humidity, 45 minutes at 75° C.(167° F.), and 135 minutes at 150° C. (302° F.) to yield a siliconemembrane 113 about 25 μm thick, and having an integrated spacer 110,whose thickness is not inclusive of the resulting silicone membrane. Thepolyester mesh fabric is adhered to the cured silicone membrane 113, butwas not totally encapsulated by the silicone membrane 113. The onesurface of the membrane 113 has a matte finish suitable for contact withblood or blood products.

Additional integrated silicone membranes having thicknesses of about 13μm and about 50 μm are fabricated using the silicone dispersion method.

Example 3 Fabrication of an Inner Collapsible Blood Container 102

A silicone blood bag is fabricated from a pair of silicone sheets bybonding the edges together with Smooth On Sil-Poxy RTV adhesive(Smooth-On, Inc. Easton, Pa.) and placing the bonded sheets between apair of flat aluminum plates to yield a silicone blood bag. A siliconeinlet tube (McMaster Carr #5236K83, McMaster Carr, Inc., Robbinsville,N.J.) is bonded within the seam to provide for fluid passage and nestedwithin a groove in the aluminum plates before clamping the platestogether with large binder clamps and allowing the adhesive to cureovernight. The silicone blood bag is removed from the aluminum platesthe next day and leak tested by insufflating with compressed air andsubmerging in water to observe for bubbles before use. The siliconeblood bag is then placed in an outer barrier bag fabricated as describedin Example 1.

The silicone blood bag is placed inside the barrier bag as disclosed inExample 1 and the silicone inlet tube of the silicone blood bag isconnected to the multilayer tube using a plastic barb fitting (McMasterCarr #5116K18, McMaster Carr, Inc., Robbinsville, N.J.), and an oxygensensor tab (Mocon #050-979, Mocon, Inc., Minneapolis, Minn.) is affixedto the inside of the barrier bag. A pair of plastic mesh spacers(McMaster Carr #9314T29, NJ McMaster Carr, Inc., Robbinsville, N.J.) arecut to about 12.7×17.8 cm (5×7 inches) and one or more sachets of oxygensorbent (Mitsubishi Gas Chemical America, New York, N.Y.) are affixednear the center of each piece of plastic mesh just seconds prior toplacing the plastic mesh spacers between the blood bag and barrier bagand sealing the final edge of the barrier bag with the impulse sealer.The resulting oxygen depletion device 10 is used in subsequent testing.

Example 4 Blood Preparation

Whole blood and blood products including leukoreduced whole blood andleukoreduced packed red blood cells are prepared using techniques knownin the art. Samples are analyzed as indicated using a Radiometer ABL-90hemoanalyzer (Radiometer America, Brea, Calif.) according tomanufacturer instructions including pH, blood gas, electrolyte,metabolite, oximetry, and baseline sO₂, and pO2 levels. Free hemoglobinis measured using the Hemocue® Plasma Low Hb Photometer according tomanufacturer instructions.

As appropriate, blood sO₂ levels are increased to levels typical ofcollected whole blood (65 to 90%) by passing the blood or bloodcomponent through a Sorin D100 oxygenator (Arvada, Colo.) with oxygen asthe exchange gas. All experiments begin with ≥50% sO₂ prior totransferring the blood to an oxygen depletion device for testing.

Example 5 Test of Deoxygenation

An oxygen depletion device of Example 2 is provided with blood andtested as follows. Whole blood (124 grams) is obtained and saturatedwith oxygen by injecting several cc of pure oxygen gas and placed in thesilicone blood bag of Example 2 by sterile transfer using a TerumoSterile Connection Device (SCD) and weighing the bag during transfer.The outer receptacle 101 headspace oxygen level is measured using aMocon OpTech Platinum oxygen analyzer and determined to be 1.60 torr atthe start of the experiment. An initial sample of blood is taken andmeasured on a Radiometer ABL-90 hemoanalyzer (Radiometer America, Brea,Calif.) and the saturated oxygen content (sO2) found to be 98.7%. Thebarrier bag with blood is placed on a work bench at room temperature(21.0° C.) and allowed to stand one hour without agitation. After onehour, the sO2 is determined to be 93.5% sO₂ and the barrier bagheadspace oxygen is determined to be 0.70 torr oxygen. The barrier bagwith blood is incubated at room temperature (21.0° C.) for about 14hours without agitation. After 14 hours incubation, the sO₂ isdetermined to be 66.7%, and a final determination of sO₂ is 51.2% afteran additional 7 hours incubation at 21° C. without agitation. The rateof deoxygenation follows first order kinetics and the rate constant iscalculated to be on the order of about min⁻¹.

Example 6 Serpentine Urethane Flow Oxygen Depletion Devices

A collapsible blood bag is fabricated from breathable polyurethane film(American Polyfilm, Branford, Conn.) having a reported moisture vaportransmission rate of 1800 gr/m²/24 hrs., wherein a serpentine tortuousflow path is fabricated using a custom heat sealing die to weld a pairof the films together to create the geometry. The collapsible bag withtortuous path comprised a series of 12 channels of about 5 mm width and220 mm in length, providing for an overall flow path of about 2640 mm.The collapsible bag is sealed within an outer barrier according toExample 1. The resulting depletion device further includes twomultilayer tubes sealed within one end, as previously described in thisdisclosure, such that the inlet and outlet of the tortuous path are influid connectivity with the pieces of multilayer tube.

Two pieces of plastic spacer mesh (McMaster Carr #9314T29, McMasterCarr, Inc., Robbinsville, N.J.) are cut to about 125×180 mm (5×7 inches)and placed on both sides of the collapsible blood container within theouter barrier receptacle. A sachet of oxygen sorbent (SS-200, MitsubishiGas Chemical America, NY, N.Y.) is placed between each of the plasticmesh spacers and the outer barrier receptacle (2 sachets total) and anoxygen sensor tab (Mocon #050-979, Mocon, Inc. Minneapolis, Minn.)before sealing the final edge of the outer barrier receptacle. A lengthof standard IV tubing (Qosina T4306, Qosina, Corp., Edgewood, N.Y.) 914mm (36 inches) is solvent bonded using cyclohexanone to each of themultilayer tubes. A ratchet clamp (Qosina #140072, Qosina, Corp.,Edgewood, N.Y.) is placed onto the outlet tubing to control flow.

A standard 500-mL blood bag (model KS-500, KS Mfg., Avon, Mass.) isconnected to the length of outlet tubing using a Terumo sterile tubingwelder (model TSCD-II, Terumo BCT, Inc., Lakewood, Colo.). A secondstandard 500-mL blood bag (model KS-500, KS Mfg., Avon, Mass.) is filledwith 325 grams of blood at 20.8° C. and a sample measured on aRadiometer ABL-90 hemoanalyzer (Radiometer America, Brea, Calif.) andfound to have 83.0% sO₂ and 70.1 mm Hg pO₂. A ratchet clamp (Qosina#140072, Qosina, Corp., Edgewood, N.Y.) is placed onto the inlet tubingto control flow and the filled blood bag is then connected to the inlettube using a Terumo sterile tubing welder (model TSCD-II, Terumo BCT,Inc., Lakewood, Colo.). The ratchet clamps are closed to prevent flowand the filled blood bag is hung from an IV pole such that the inlettubing was fully extended and the collapsible blood bag is on thelaboratory bench. The outlet bag is tared on a balance before it isplaced on the floor with the outlet tubing fully extended. The headspaceoxygen level in the outer receptacle is measured using a Mocon Op-Techplatinum oxygen analyzer and found to be 0.05 torr oxygen at the start.The clamps are opened and a stopwatch timer started to measure theduration of flow, and after 3 minutes 25 seconds the inlet blood bag isemptied and the ratchet clamps are closed. A sample of blood is takenand measured and found to have 84.1% sO₂ and 71.6 mmHg pO₂, the increasepresumably from residual oxygen in the empty circuit. The headspacemeasured 0.00 torr oxygen and the outlet blood bag contained 277 gramsof blood.

The empty inlet blood bag is removed from the IV pole and placed on thefloor, while the outlet blood bag with 277 grams of blood is hung fromthe IV pole to repeat the flow. The IV pole is lowered to 457 mm (18inches) to reduce the flow rate and the clamps opened to repeat thecycle. The process is repeated 5 times, and then the collapsible bloodbag is filled with blood and allowed to remain stationary on thelaboratory bench for 80 minutes and a terminal blood sample is taken formeasurements on the hemoanalyzer. The table below summarizes theresults, which show a slight gradual increase in the oxygen level of theblood during flow, with a slight decrease after standing. The resultsindicate that the system does not provide for an appreciabledeoxygenation of the blood over the course of the study, but ratherabsorbs oxygen from the permeable standard PVC blood bags. From this,the importance of taking additional measures to prevent the ingress ofoxygen at inlets, outlets, ports, and tubing is demonstrated.

TABLE 3 Deoxygenation using urethane bags Flow Pass # Time (min:sec) sO₂% pO₂ mmHg Headspace O₂ torr Start n/a 83.0 70.1 0.05  1* 3:25 84.1 71.60.00 2 7:45 84.8 72.6 0.09 3 6:38 85.2 72.9 0.09 4 7:16 85.6 73.4 0.13 56:31 85.4 72.9 0.15 Stagnant 80:00  83.6 64.3 0.15 *914 mm head height;all other flow passes at 457 mm height.

Example 7 Test of Inner Collapsible Blood Container 102 Configurations

A series of inner collapsible blood containers 102 are preparedaccording to Table 4 below and sealed in an outer receptacle 101 asprovided in Example 1. Leukoreduced packed red blood cells (LRpRBC) areintroduced into the container 102. The resulting oxygen depletiondevices 10 further included a Mocon Optech-O2 sensor. Assembled bloodcontainers according to Table 4 are placed on a Helmer Labs PlateletShaker, Model PF96 and blood and headspace samples are obtained andanalyzed at time points between 0 and 300 minutes.

TABLE 4 Inner collapsible blood container 102 test configurationsThickness # LRpRBC or Pore Sorbent Sample Hct sO_(2i) Material Sourcesize Compartments Sachets* 1A >45% >80% Silicone Wacker 30 μm Double 10Silpuran 2A >45% >80% Silicone McMaster- 127 μm  Single 2 Carr3A >45% >80% Silicone Polymer 76 μm Triple 15 Science 4A >45% >80%Silicone Polymer 76 μm Triple 15 Science 5A >45% >80% PVDF Millipore0.22 μm   Singe 5 GVSP 1B 52% >50% PVDF Millipore 0.22 μm   Single 5VVSP 2B 52% >50% PVDF Millipore 0.1 μm  Single 5 VVSP 3B 52% >50% PVDFMillipore 1.0 μm  Single 5 VVSP 4B 52% >50% Silicone Wacker 50 μm Single5 5B 52% >50% Silicone Wacker 20 μm Single 5 6B 52% >50% SiliconePolymer 76 μm Single Science All oxygen depletion devices include anouter receptacle 101 according to Example 1. All examples incorporate aspacer 110 to maintain headspace

As shown in FIG. 5, the depletion of oxygen follows first orderkinetics. The rate constants are provided in Table 5.

TABLE 5 Rate constants Sample Rate Constant (min⁻¹) Simulation −1.20 ×10⁻² 1A −1.00 × 10⁻² 2A −0.41 × 10⁻² 3A −0.62 × 10⁻² 4A −0.82 × 10⁻² 5A−0.93 × 10⁻² 1B −1.12 × 10⁻² 2B n/a 3B −1.40 × 10⁻² 4B −1.03 × 10⁻² 5B−1.34 × 10⁻² 6B −0.95 × 10⁻²

Example 8 30.5×30.5 cm (12×12 Inch) Thick Silicone Bag

A collapsible blood container 102 is fabricated from a pair of siliconesheets 152 μm thick and 228 μm thick, respectively (McMaster Carr#87315K71, McMaster Carr, Inc., Robbinsville, N.J.), bonded togetheraround the periphery Sil-Poxy silicone adhesive (Smooth-On, Inc.,Easton, Pa.) and bonding silicone tubing (McMaster Carr #9628T42,McMaster Carr, Inc., Robbinsville, N.J.) for fluid communication as aninlet tube. The bonded sheets are cured two days between clampedaluminum plates.

The collapsible blood bag inlet tube is connected with a multilayer tubeof an outer receptacle barrier bag 101 according to Example 1 with aplastic barb fitting (McMaster Carr #5116K18, McMaster Carr, Inc.,Robbinsville, N.J.). The resulting outer receptacle bag 101 is leaktested by insufflation submersion as described in Example 1.

The device 10 is assembled with two 330×330 mm mesh spacers (McMasterCarr #9314T29, McMaster Carr, Inc., Robbinsville, N.J.) and four sachetsof oxygen sorbent are affixed to each mesh spacer with tape (SS-200,Mitsubishi Gas Chemical America, NY, N.Y.), the collapsible bloodcontainer, and an oxygen sensor tab (Mocon #050-979, Mocon, Inc.Minneapolis, Minn.) inserted and heat sealed in the barrier bag 101. Theinlet tube comprised standard IV tubing (200 mm Qosina T4306, Qosina,Corp., Edgewood, N.Y.) is solvent bonded with cyclohexanone to themultilayer tube of barrier bag 101. A ratchet clamp (Qosina #140072,Qosina, Corp., Edgewood, N.Y.) provides for control flow.

A pair of matched units of blood are prepared for the study by adjustingthe hematocrit to 50% after centrifugation and recombining the red cellsand desired amount of plasma to achieve the target hematocrit. Theinitial blood sO₂ is measured on a Radiometer ABL-90 hemoanalyzer(Radiometer America, Brea, Calif.) and found to have 39.0% sO₂; threesyringes containing 30-cc of 100% oxygen were added to the blood toobtain a sO₂ level of 85.6% before starting the study. A portion of theblood is transferred to a tared standard 500-mL blood bag (model KS-500,KS Mfg., Avon, Mass.) and filled with 532 grams of blood to represent atypical 500-mL unit of donated blood having a high hematocrit andsaturated oxygen level. The standard 500-mL blood bag (model KS-500, KSMfg., Avon, Mass.) is connected to the length of inlet tubing using aTerumo sterile tubing welder (model TSCD-II, Terumo BCT, Inc., Lakewood,Colo.) and the contents transferred into the collapsible blood bag undertest.

The collapsible blood bag is placed on a Helmer model PF96 plateletagitator (Helmer Scientific, Nobelsville, Ind.) and a sample of blood istaken and measured on a Radiometer ABL-90 hemoanalyzer (RadiometerAmerica, Brea, Calif.) and the headspace oxygen level measured on aMocon Op-Tech platinum oxygen analyzer (Mocon, Inc., Minneapolis,Minn.). At the beginning, the blood is found to have 84.5% sO₂ and theheadspace oxygen partial pressure is 2.48 torr. Samples are agitated onthe platelet agitator and samples taken and measured every 30 minutesfor 150 minutes duration. The results are summarized in Table 6 below.

TABLE 6 Deoxygenation using a 30.5 × 30.5 cm silicon bag Headspace O₂Time (min) sO₂ % pO₂ mmHg pCO₂ mmHg torr 0 84.5 63.9 88.9 2.48 30 70.340.0 50.7 4.63 60 64.9 34.9 43.6 5.24

The calculated deoxygenation rate constant is −0.34×10⁻² min⁻¹.

Example 9 Effect of Mixing on Oxygen Depletion

Four oxygen reduction bags (ORB), having a silicone inner collapsibleblood container 102, are prepared according to Example 3. The innercollapsible blood container 102, is filled with leukocyte reduced packedred blood cells (LRPRBC), prepared according to Example 4, to obtain asurface area to volume (SAV) ratio of approximately 6 cm²/ml. ThreeLRPRBC filled ORBs are placed flat on a Helmer PF-96 Platelet Agitator(Noblesville, Ind.) or a PF-8 agitator, with the standard cycles perminute (72 cpm) or modified to a reduced-standard cpm (42 cpm) linearoscillation. A third set of filled ORB's are placed on a Benchmark 3D5RVH6 agitator (Sayretville, N.J.). Samples are collected and analyzedat 0, 60, 120, and 180 mins for various ABL-90 outputs outlined inExample 4.

As shown in FIG. 12, 3D mixing results in the highest rate of oxygendepletion and the lowest percent sO₂ at T₁₈₀ compared to the linearoscillation method of mixing. Further, a higher rate of oxygen depletionis obtained with the standard cpm linear oscillation (SLO) compared tothe reduced-standard cpm linear oscillation (R-SLO).

Example 10 Effect of Surface to Volume Ratios on Oxygen Depletion

In another example, six oxygen reduction bags (ORB), having an innercollapsible blood container 102, are prepared with Bentec silicone.LRPRBC is collected and prepared according to Example 4. The siliconeinner collapsible blood containers 102, are filled with 176, 220, 250,270, 300, and 350 mL of LRPRBC, to provide the surface to area ratiosbetween 3.41-6.8 cm²/ml as shown in Table 7. The percent sO2 is measuredin the ORBs containing the various LRPRBC volumes at 0, 30, 60, 120, and180 mins as described in Example 4.

TABLE 7 Surface Area to Volume Ratio Blood Volume (ml) 176 220 250 270300 350 SAV Ratio (cm²/ml) 6.8 5.45 4.8 4.4 4 3.41

As shown in FIG. 13, the surface area kinetic rates decrease once theSAV ratio is below 5.45 cm²/ml.

In another example, five oxygen reduction bags (ORB), having an innercollapsible blood container 102, are prepared with PVDF instead ofsilicone. LRPRBC is collected and prepared according to Example 4. ThePVDF inner collapsible blood containers 102 are filled with 95, 110,220, 300, or 360 ml blood volume, to provide the surface area to volumeratios shown in Table 8. The percent sO2 is measured in the ORBscontaining the various LRPRBC volumes at 0, 30, 60, 120, and 180 mins asdescribed in Example 4.

As shown in FIG. 14, the lowest percent sO2 after 180 mins is achievedwhen the SAV ratio is above 5.

TABLE 8 Surface Area to Volume Ratio Blood Volume (ml) 95 110 220 300360 SAV Ratio (cm²/ml) 6.8 5.9 2.9 2.2 1.8 Kinetic Rate (×100) −1.39−1.9 −0.82 −0.06 −0.32

In another example, four oxygen reduction bags (ORB), having single ordouble sided membranes (PSU or PVDF) are prepared. LRPRBC is collectedand prepared according to Example 4. The inner collapsible bloodcontainers 102 are filled with 112-118 ml LRPRBC, to provide a 50%reduction in surface area volume in the double sided membrane. Thepercent sO2 is measured in the ORBs containing the single or doublemembrane as described in Example 4. As shown in FIG. 15, a 50% reductionin surface area results in a 50-60% reduction in the overall kineticrate.

Example 11 Preparation of Collapsible Blood Containers from MicroporousPolysulfone or PVDF

Heat sealed polysulfone and PVDF oxygen permeable collapsible bloodcontainers 102 are prepared. The prepared seal results in the breakdownof the microporous structure of the films to produce a crystalline areathat is sensitive to the flexural stresses associated with fluidmovement in the resulting containers 102. The containers 102 are subjectto leakage and breakage and are not suitable for ORB's intended for useoutside of an experimental setting.

To overcome the inability to heat seal polysulfone or PVDF membranes ina manner suitable for use in transfusion medicine, a heat laminable“tie” layer 105 is included in the construction of the containers 102prepared from microporous membranes 113. As shown in FIG. 9B, the sealarea is reinforced by pre-laminating low density polyethylene (LDPE)strips to the inside surfaces of the upper and lower membranes to alignwith the bag seal area. The pre-lamination results in pre-laminate seals107 as shown in FIG. 9B. The two pre-laminated membranes 113 (114) arethen heat sealed to form the seal 108 as shown in FIG. 9B. Also as shownin FIG. 9B, the tie layers extend beyond the width of the seal by anamount.

LDPE melts at about 105° C., well below the melting temperatures ofpolysulfone (187° C.) or PVDF (177° C.). The bag is completed byaligning the seal areas and heat sealing the upper and lower membranestogether to form a bag. Without being limited to a specific mechanism,it is believed that the LDPE flows into the membrane pores, acting as areinforcing strain relief inboard of the seal and a low temperature“tie” layer for the seal.

In addition to strengthening the seal between the microporous membranes113, the tie layer also serves as a geometric feature 121. Thus theoverall internal geometry can be readily adjusted by selecting the shapeof the tie layers 105. Examples of exemplary geometries are shown inFIG. 10. As shown, the geometric feature 121 provides a rounded internalgeometry thereby avoiding reduced mixing associated with corners. Asshown in FIG. 10, the resulting container 102 can be oval or round andmay further include a mixing feature 119 which provides for a circularflow of blood product and enhances mixing.

An inner collapsible blood bag is fabricated from a pair of MilliporePVDF membranes having 0.22 μm pore size, 177×177 mm square, by firstheat bonding a low density polyethylene (LDPE) tie layer frame to eachmembrane. The LDPE tie layer frame has a thickness of about 0.02-0.10 mmand is about 177×177 mm square on the outside dimensions and has aninside dimension that is about 160×160 mm square for use with a 15 mmwide seal, thereby providing for about 4 mm of overlap between the edgeof the seal and the end of the tie layer to provide stress relief in theseal edge. The tie layer frame is heat bonded to the PVDF membrane usingan impulse heat sealer. The pair of tie layer bonded membranes are thenheat sealed together around their periphery using a pair of customfabricated constant heat aluminum dies having a tube sealing groove, aspreviously described, to yield an inner collapsible blood container. Apair of Conwed Thermanet part #R03470 polymer integrating mesh sheetsare cut to approximately 10 mm larger than the inner collapsible bloodbag periphery and placed on both sides of the inner collapsible bloodbag with the adhesive sides of the polymer integrating mesh in contactwith the inner collapsible blood bag. The assembly is placed between apair of aluminum plates and heated to about 93-110° C., up to as much as120° C., for about 3-15 minutes to melt the adhesive and allow it toflow into the pores of the PVDF membrane directly underneath theintegrating polymer mesh, and also around the periphery of the innercollapsible blood bag where the polymer integrating mesh is in contactwith itself, thereby providing a strong mechanical bond. The assembly isallowed to cool below about 50° C. before removing the plates.

Example 12 Effect of Spacer 110 on Deoxygenation Rates

Inner collapsible blood bag and oxygen depletion device are accordingthe examples above with and without a spacer 110 according to thepresent disclosure. As shown in FIG. 16, incorporation of a spacer 110significantly increases the rate of oxygen depletion.

While the invention has been described with reference to particularembodiments, it will be understood by those skilled in the art thatvarious changes may be made and equivalents may be substituted forelements thereof without departing from the scope of the invention. Inaddition, many modifications may be made to adapt a particular situationor material to the teachings of the invention without departing from thescope of the invention.

Therefore, it is intended that the invention not be limited to theparticular embodiments disclosed as the best mode contemplated forcarrying out this invention, but that the invention will include allembodiments falling within the scope and spirit of the appended claims.

The invention claimed is:
 1. An oxygen depletion device 10 for depletingoxygen from blood comprising: an outer receptacle 101 substantiallyimpermeable to oxygen; an inner collapsible blood container 102comprising one or more chambers comprising polyvinyl chloride (PVC) orpolyolefin; a spacer 110 situated between said outer receptacle 101 andsaid inner collapsible blood container 102; and an oxygen sorbent 103situated between said outer receptacle 101 and said inner collapsibleblood container 102, wherein said spacer 110 is a mesh comprising atleast one interstice 111 and maintains a headspace defined by said outerreceptacle 101 and said inner collapsible blood container 102, whereinsaid at least one interstice 111 comprises between 50% to 80% of thetotal area of said mesh material of said spacer 110, and wherein saidheadspace ensures efficient diffusion of oxygen from a surface of saidinner collapsible blood container 102 to said oxygen sorbent
 103. 2. Theoxygen depletion device 10 of claim 1, wherein said inner collapsibleblood container 102 comprises a multilayered membrane.
 3. The oxygendepletion device 10 of claim 1, wherein said inner collapsible bloodcontainer 102 comprises PVC comprising a citrate plasticizer.
 4. Theoxygen depletion device 10 of claim 1, wherein said inner collapsibleblood container 102 comprises a multilayered membrane comprising atleast one layer consisting of polyester.
 5. The oxygen depletion device10 of claim 1, wherein said inner collapsible blood container 102comprises PVC comprising DEHP plasticizer.
 6. The oxygen depletiondevice 10 of claim 1, wherein said oxygen sorbent 103 further comprisesa carbon dioxide sorbent
 103. 7. The oxygen depletion device 10 of claim6, wherein said oxygen and carbon dioxide sorbent 103 is situated withina sachet disposed within said headspace.
 8. The oxygen depletion device10 of claim 1, wherein said inner blood compatible container 102 has asurface area to volume ratio of at least 4.48 centimeters/milliliter(cm²/ml) when filled with a unit of blood for depletion and enclosedwithin said outer receptacle
 101. 9. The oxygen depletion device 10 ofclaim 1, wherein said inner collapsible blood container 102 comprises afirst and second membrane 113 joined by a peripheral tie layer
 105. 10.The oxygen depletion device 10 of claim 9, wherein said peripheral tielayer 105 consists of silicone.
 11. The oxygen depletion device 10 ofclaim 1, wherein said headspace has a volume of between 10 and 500milliliters (ml).
 12. The oxygen depletion device 10 of claim 1, whereinsaid at least one interstice 111 comprises between 60% to 75% of thetotal area of said mesh material of said spacer
 110. 13. The oxygendepletion device 10 of claim 1, further comprising at least oneinlet/outlet 30 passing through said outer receptacle 101 comprising atube 301, a bond 302, and a port 303, wherein said tube 301 and saidbond 302 are substantially impermeable to oxygen, and said inlet/outlet30 is in fluid communication with said inner collapsible blood container102.
 14. The oxygen depletion device 10 of claim 1, wherein said innercollapsible blood container 102 comprises a mixing structure
 119. 15.The oxygen depletion device 10 of claim 1, wherein blood collected insaid oxygen depletion device 10 is deoxygenated at a rate constant of atleast −0.5×10⁻² min⁻¹.
 16. The oxygen depletion device 10 of claim 1,wherein said inner collapsible blood container 102 comprises ahydrophobic material.
 17. The oxygen depletion device 10 of claim 1,wherein said inner collapsible blood container 102 does not comprise aplasticizer.
 18. The oxygen depletion device 10 of claim 1, wherein saidat least one interstice 111 has an open area of greater than 0.75 squaremillimeters (mm²).